CN103617375B - The method and system of PCR product sequencing and typing - Google Patents

Abstract

The present invention proposes a method for sequencing and typing polymerase chain reaction products, comprising the following steps: providing base sequences to be typed, file information and primer file information; allele type information based on file information and reference sequences Identify the alignment positional relationship between the base sequence to be typed and the reference sequence, and obtain the first candidate type combination to be analyzed; perform typing and recognition on the base sequence to be typed according to the primer file information and the primer information of the reference sequence, and obtain the second A combination of candidate types to be analyzed; the final type of the base sequence to be typed is obtained according to the first and second combination of candidate types to be analyzed. The embodiment according to the present invention has the advantages of high typing efficiency, accurate and reliable typing, short typing time and low labor cost. The invention also provides a system for sequencing and typing polymerase chain reaction products.

Description

Method and system for sequencing and typing polymerase chain reaction products

technical field

The invention relates to the technical field of allele typing, in particular to a method and system for sequencing and typing polymerase chain reaction products.

Background technique

HLA (Human Leucocyte antigen), that is, human leukocyte antigen. It is a group of glycoproteins encoded by the major histocompatibility complex (MHC) gene. For the human immune system, HLA and MHC are the same concept, and HLA is also the human major compatibility complex . HLA typing refers to the detection of individual HLA antigen-specific genes by means of serological, cytological or molecular biological methods. In clinical organ transplantation, HLA typing is the main basis for selecting suitable donors.

The current international standard HLA typing techniques are PCR-SSP (Polymerase Chain Reaction Sequence-Specific Primers, sequence-specific primer polymerase chain reaction), PCR-SSO (Polymerase Chain Reaction sequence-Specific Oligonucleotide Probe Hybridization, polymerase chain reaction oligonucleotide Nucleic acid probe hybridization) and PCR-SBT (Polymerase Chain Reaction Sequencing-based Typing, polymerase chain reaction product direct sequencing typing).

PCR-SBT uses primers to amplify the polymorphic regions of the HLA gene by PCR, then performs DNA sequencing on the amplified products, and determines the HLA allele type with the assistance of computer software. For the analysis of gene structure, SBT is a relatively intuitive and accurate method, and can directly discover new alleles. Novel alleles are identified using PCR-SSP or PCR-SSO methods and are usually also confirmed by sequencing.

In SBT technology, after the DNA is sequenced using the amplified product, it is necessary to use software to compare the sequencing results with the HLA standard sequence published in the international organization IMGT database (http://www.ebi.ac.uk/imgt/) ; Obtain the matching rate of the sample sequence and the standard sequence through comparison; draw the typing conclusion of the sample sequence according to the matching rate.

Currently, relatively successful typing software include uType and SBTengine, both of which are based on PCR-SBT sequencing.

However, as the number of alleles in the IMGT database increases, the slower the speed and the lower the efficiency when performing a large-scale candidate type screening search during the typing process of the above software. In addition, in the manual-assisted typing stage, when the typing personnel check the electropherogram, the operation procedure is cumbersome and takes a long time. At the same time, the existing HLA typing software is only applicable to Windows systems, but cannot run on Linux and Mac systems.

Contents of the invention

The purpose of the present invention is to solve at least one of the above-mentioned technical drawbacks.

Therefore, the object of the present invention is to propose a method for sequencing and typing polymerase chain reaction products. The method has the advantages of high typing efficiency, accurate and reliable typing, short typing time and low labor cost.

Another object of the present invention is to provide a system for sequencing and typing polymerase chain reaction products.

In order to achieve the above purpose, an embodiment of the present invention provides a method for sequencing and typing polymerase chain reaction products, including the following steps: obtaining the base sequence to be typed, and the base sequence associated with the base sequence to be typed File information and primer file information, wherein the file information includes the site name of the base sequence to be typed; the base to be typed is identified according to the file information and the allele type information of the reference sequence sequence and the alignment positional relationship of the reference sequence to obtain the first candidate type combination to be analyzed; according to the primer file information and the primer information of the reference sequence, the base sequence to be typed is further typed and identified, to obtain a second combination of candidate types to be analyzed; and to obtain the final type of the base sequence to be typed according to the first combination of candidate types to be analyzed and the second combination of candidate types to be analyzed.

According to the method for sequencing and typing of polymerase chain reaction products according to the embodiment of the present invention, the link between the base sequence to be typed and the reference sequence can be directly identified according to the reference sequence and its related information and the base sequence to be typed and its related information. Therefore, the step of aligning the read base sequence to the corresponding position of each reference sequence is reduced, and the comparison time is shortened. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened. In addition, the re-identification and typing are carried out according to the primer information, thus ensuring the correctness and reliability of the typing. In addition, the method can automatically type the base sequence to be typed, reducing the tediousness of manually importing GSSP information, effectively saving time and reducing labor costs.

In addition, the method for sequencing and typing polymerase chain reaction products according to the above-mentioned embodiments of the present invention may also have the following additional technical features:

In some examples, the alignment position relationship between the base sequence to be typed and the reference sequence is identified according to the file information and the allelic type information of the reference sequence, so as to obtain the first candidate type combination to be analyzed , further comprising: matching the site name of the base sequence to be typed included in the file information with the allelic type information of the reference sequence; obtaining the base sequence to be typed according to the matching result The alignment positional relationship with the reference sequence; according to the alignment positional relationship, the first combination of candidate types to be analyzed is obtained.

Further, the allelic type information of the reference sequence includes the HLA type I and type II site gene information of IMGT, and the site name of the base sequence to be typed included in the file information is the corresponding site locus gene information.

In some examples, further typing and identifying the base sequence to be typed according to the primer file information and the primer information of the reference sequence to obtain a second candidate type combination to be analyzed further includes: Matching the primer file information and the primer information of the reference sequence; if the matching is consistent, adding the corresponding reference sequence to the second candidate type combination to be analyzed.

Further, the primer information of the reference sequence includes GSSP primer information of HLA type I and II, that is, backup primer information.

In some examples, the obtaining the final type of the base sequence to be typed according to the first candidate type combination to be analyzed and the second candidate type combination to be analyzed further includes: obtaining the first An intersection of the combination of candidate types to be analyzed and the second combination of candidate types to be analyzed; according to the intersection, the final type of the base sequence to be typed is determined.

The embodiment of the second aspect of the present invention provides a system for sequencing and typing polymerase chain reaction products, including: a storage module for storing the base sequence to be typed, and the base sequence associated with the base sequence to be typed File information and primer file information, as well as reference sequence, allele type information and primer information of the reference sequence, wherein the file information includes the site name of the base sequence to be typed; the analysis module uses Identifying the alignment positional relationship between the base sequence to be typed and the reference sequence based on the file information and the allelic type information of the reference sequence to obtain the first candidate type combination to be analyzed, and according to the primer The file information and the primer information of the reference sequence are further typed and identified on the base sequence to be typed to obtain a second candidate type combination to be analyzed, and according to the first candidate type combination to be analyzed and the Combining the second candidate types to be analyzed to obtain the final type of the base sequence to be typed; and a display module for displaying the information stored in the storage module and the analysis results of the analysis module.

According to the polymerase chain reaction product sequencing and typing system of the embodiment of the present invention, according to the reference sequence and its related information, the base sequence to be typed and its related information can directly identify the combination of the base sequence to be typed and the reference sequence. Therefore, the step of aligning the read base sequence to the corresponding position of each reference sequence is reduced, and the comparison time is shortened. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened. In addition, the re-identification and typing are carried out according to the primer information, thus ensuring the correctness and reliability of the typing. In addition, the system can automatically type the base sequence to be typed, reducing the tediousness of manually importing GSSP information, effectively saving time and reducing labor costs.

In addition, the system for sequencing and typing polymerase chain reaction products according to the above embodiments of the present invention may also have the following additional technical features:

In some examples, the analysis module is used to: match the site name of the base sequence to be typed included in the file information with the allele type information of the reference sequence; obtain The alignment positional relationship between the base sequence to be typed and the reference sequence; the first candidate type combination to be analyzed is obtained according to the alignment positional relationship.

Further, the allelic type information of the reference sequence includes the HLA type I and type II site gene information of IMGT, and the site name of the base sequence to be typed included in the file information is the corresponding site locus gene information.

In some examples, the analysis module is also used to: match the primer file information with the primer information of the reference sequence; if the match is consistent, add the corresponding reference sequence to the second candidate type to be analyzed Do not combine.

Further, the primer information of the reference sequence includes GSSP primer information of HLA type I and II, that is, backup primer information.

In some examples, the analysis module is also used to: obtain the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed; determine the base to be typed according to the intersection The final type of the sequence.

Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.

Description of drawings

The stated and/or additional aspects and advantages of the present invention will become apparent and easily understood from the following description of the embodiments in conjunction with the accompanying drawings, wherein:

1 is a flowchart of a method for sequencing and typing polymerase chain reaction products according to an embodiment of the present invention;

2 is a schematic diagram of resource consumption for typing different numbers of reference sequences (samples) according to a method for sequencing and typing polymerase chain reaction products according to an embodiment of the present invention;

Fig. 3 is the operation flowchart of typing processing by applying the method for sequencing and typing polymerase chain reaction products according to one embodiment of the present invention;

Fig. 4 is a schematic diagram of the operation interface of the method for sequencing and typing polymerase chain reaction products using an embodiment of the present invention; and

Fig. 5 is a structural block diagram of a system for sequencing and typing polymerase chain reaction products according to an embodiment of the present invention.

detailed description

Embodiments of the present invention are described in detail below, examples of which are shown in the drawings, wherein the same or similar reference numerals designate the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by referring to the figures are exemplary only for explaining the present invention and should not be construed as limiting the present invention.

In describing the present invention, it should be understood that the terms "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", The orientation or positional relationship indicated by "horizontal", "top", "bottom", "inner", "outer", etc. are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than Nothing indicating or implying that a referenced device or element must have a particular orientation, be constructed, and operate in a particular orientation should therefore not be construed as limiting the invention.

In the description of the present invention, it should be noted that unless otherwise specified and limited, the terms "installation", "connection" and "connection" should be understood in a broad sense, for example, it can be a mechanical connection or an electrical connection, or it can be two The internal communication of each element may be directly connected or indirectly connected through an intermediary, and those skilled in the art can understand the specific meaning of the terms according to specific situations.

The method and system for sequencing and typing polymerase chain reaction products according to the embodiments of the present invention will be described below with reference to the accompanying drawings.

Fig. 1 is a flowchart of a method for sequencing and typing polymerase chain reaction products according to an embodiment of the present invention. As shown in Figure 1, the method for sequencing and typing polymerase chain reaction products according to one embodiment of the present invention includes the following steps:

Step S101: Obtain the base sequence to be typed, the file information associated with the base sequence to be typed, and the primer file information, wherein the file information includes the site name of the base sequence to be typed.

Specifically, the base sequence to be typed, the file information associated with the base sequence to be typed and the primer file information may be pre-stored in the database.

For example: the base sequence to be typed, the file information associated with the base sequence to be typed, and the primer file information are stored in a dynamic database, which is mainly used to store and analyze samples (ie, the base sequence to be typed) Intermediate result data in . The dynamic database stores, for example, three database tables, namely a file information table, a primer file information table, and a sample information table. The file information table stores the useful information of each sequence file (for example: the file information associated with the base sequence to be typed); the primer file information table stores the useful information of each primer file (for example: the file information associated with the base sequence to be typed primer file information); the sample information table stores the useful information of each sample (for example: the base sequence to be typed).

Step S102: Identify the alignment positional relationship between the base sequence to be typed and the reference sequence according to the file information and the allelic type information of the reference sequence, so as to obtain the first candidate type combination to be analyzed.

In one embodiment of the present invention, it can be realized through the following steps:

Step S1021: Match the site name of the base sequence to be typed with the allelic type information of the reference sequence according to the file information.

Step S1022: Obtain the alignment positional relationship between the base sequence to be typed and the reference sequence according to the matching result.

Step S1023: Obtain the first combination of candidate types to be analyzed according to the alignment positional relationship.

Wherein, the allelic type information of the reference sequence includes IMGT HLA type I and type II site gene information, and the site name of the base sequence to be typed included in the file information is the site gene information of the corresponding site.

Specifically, the reference sequence and its associated information can be stored in a pre-set static database, which also stores three database tables, namely the gene information table (that is, the gene information storing the reference sequence), the type Information table (that is, store the type of the reference sequence) and primer information table (that is, store the primer information of the reference sequence). More specifically, the gene information table stores the gene information of HLA type I and type II loci from IMGT, the type information table stores all allele type information, and the primer information table stores HLA type I, type II All GSSP primers and alternate primer information.

In this way, the base sequence to be typed can be preliminarily typed through the reference sequence and its related information in the static database and the base sequence to be typed and its related information in the dynamic database. As mentioned above, for example, according to the above-mentioned information, the alignment positional relationship between the base sequence to be typed and the reference sequence can be directly identified, thereby obtaining the first combination of candidate types to be analyzed, thereby reducing the need for reading base sequences one by one. The step of aligning to the corresponding positions of each reference sequence reduces the alignment time. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened.

Step S103: Perform further typing identification on the base sequence to be typed according to the primer file information and the primer information of the reference sequence, so as to obtain the second candidate type combination to be analyzed.

In one embodiment of the present invention, the following steps may be included:

Step S1031: matching the primer file information with the primer information of the reference sequence.

Step S1032: If the matches are consistent, add the corresponding reference sequence to the second combination of candidate types to be analyzed.

Wherein, the primer information of the reference sequence includes the GSSP primer information of HLA type I and II, that is, the spare primer information.

That is to say, when identifying the base sequence of GSSP primers for sequencing, the GSSP primer information can be directly called from the static database to interpret the base sequence to be typed, so as to obtain the second candidate type combination to be analyzed. That is, by comparing the primer information associated with the base sequence to be typed with the primer information of the reference sequence, the above-mentioned second candidate type combination to be analyzed is obtained.

Step S104: Obtain the final type of the base sequence to be typed according to the first combination of candidate types to be analyzed and the second combination of candidate types to be analyzed.

Specifically, it can be achieved through the following steps:

Step S1041: Obtain the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed.

Step S1042: Determine the final type of the base sequence to be typed according to the intersection.

That is: the final type of the base sequence to be typed is confirmed according to the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed.

As shown in FIG. 2 , it is a schematic diagram of memory, processor CPU time and typing (ie analysis) time consumed for typing different numbers of reference sequences by applying the method of the embodiment of the present invention. It can be seen from Fig. 2 that even if it takes only a few minutes for the typing of 88 samples (ie base sequences to be typed), the typing can be completed.

The method in the embodiment of the present invention can be realized by software, and the software can exist in the form of a software product. In this way, when the software is manually operated for classification, it only needs to import some necessary data. Specifically, as shown in Fig. 3 and Fig. 4, wherein, Fig. 3 is a step of manual processing when applying the method of the embodiment of the present invention for classification, and Fig. 4 is a schematic diagram of an operation interface, in which , in Figure 4, 1 indicates base navigation, 2 indicates sequence display, 3 indicates electropherogram display, 4 indicates sample list, and 5 indicates type list. As shown in Fig. 3 and Fig. 4, for example, the method of the embodiment of the present invention is artificially used for classification, and only the following steps are required:

Step S301: Import files.

Step S302: Click a sample file in the file list module.

Step S303: check the mismatched sites.

Step S304: base editing.

Step S305: Check the type list module.

Step S306: According to the sequence file information, edit the mismatched bases until a type or type combination with 0 mismatches appears.

Step S307: judging whether Gssp primers are needed. If yes, execute step S308, otherwise execute step S311.

Step S308: Save.

Step S309: Running Gssp.

Step S310: import the saved file and the Gssp file, and return to step S306.

Step S311: Save.

Step S312: mark.

Step S313: Save.

Step S314: Export the message.

According to the method for sequencing and typing of polymerase chain reaction products according to the embodiment of the present invention, the link between the base sequence to be typed and the reference sequence can be directly identified according to the reference sequence and its related information and the base sequence to be typed and its related information. Therefore, the step of aligning the read base sequence to the corresponding position of each reference sequence is reduced, and the comparison time is shortened. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened. In addition, the re-identification and typing are carried out according to the primer information, thus ensuring the correctness and reliability of the typing. In addition, the method can automatically type the base sequence to be typed, reducing the tediousness of manually importing GSSP information, effectively saving time and reducing labor costs.

A further embodiment of the present invention provides a system for sequencing and typing polymerase chain reaction products. As shown in FIG. 5 , a system 500 for sequencing and typing polymerase chain reaction products according to an embodiment of the present invention includes: a storage module 510 , an analysis module 520 and a display module 530 .

Wherein, the storage module 510 is used to store the base sequence to be typed, the file information associated with the base sequence to be typed, and the primer file information, as well as the reference sequence, the allele type information of the reference sequence, and the primer information, wherein, The file information includes the site name of the base sequence to be typed.

The storage module 510 can be divided into a static database and a dynamic database. For example: the base sequence to be typed, the file information associated with the base sequence to be typed, and the primer file information are stored in a dynamic database, which is mainly used to store and analyze samples (ie, the base sequence to be typed) intermediate result data. The dynamic database stores, for example, three database tables, namely a file information table, a primer file information table, and a sample information table. The file information table stores the useful information of each sequence file (for example: the file information associated with the base sequence to be typed); the primer file information table stores the useful information of each primer file (for example: the file information associated with the base sequence to be typed primer file information); the sample information table stores the useful information of each sample (for example: the base sequence to be typed).

The reference sequence and its associated information can be stored in a static database, which also stores 3 database tables, namely the gene information table (that is, the gene information that stores the reference sequence), the type information table (that is, the gene information that stores the reference sequence type) and the primer information table (that is, the primer information that stores the reference sequence). More specifically, the gene information table stores the gene information of HLA type I and type II loci from IMGT, the type information table stores all allele type information, and the primer information table stores HLA type I, type II All GSSP primers and alternate primer information.

The analysis module 520 is used to identify the alignment positional relationship between the base sequence to be typed and the reference sequence according to the file information and the allelic type information of the reference sequence, so as to obtain the first candidate type combination to be analyzed, and according to the primer file information and the primer information of the reference sequence to further type and identify the base sequence to be typed to obtain the second candidate type combination to be analyzed, and obtain the candidate type combination according to the first candidate type combination to be analyzed and the second candidate type combination to be analyzed. The final type of the typed base sequence.

Specifically, the analysis module 520 is used to: match the site name of the base sequence to be typed included in the file information with the allelic type information of the reference sequence, and obtain the base sequence to be typed and The alignment positional relationship of the reference sequence, and obtaining the first candidate type combination to be analyzed according to the alignment positional relationship.

In this way, the base sequence to be typed can be preliminarily typed through the reference sequence and its related information in the static database and the base sequence to be typed and its related information in the dynamic database. As mentioned above, for example, according to the above-mentioned information, the alignment positional relationship between the base sequence to be typed and the reference sequence can be directly identified, thereby obtaining the first combination of candidate types to be analyzed, thereby reducing the need for reading base sequences one by one. The step of aligning to the corresponding positions of each reference sequence reduces the alignment time. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened.

In one embodiment of the present invention, the analysis module 520 is also used to: match the primer file information with the primer information of the reference sequence, and if the matching is consistent, add the corresponding reference sequence to the second candidate type combination to be analyzed . Wherein, the primer information of the reference sequence includes the GSSP primer information of HLA type I and II, that is, the spare primer information.

That is to say, when identifying the base sequence of GSSP primers for sequencing, the GSSP primer information can be directly called from the static database to interpret the base sequence to be typed, so as to obtain the second candidate type combination to be analyzed. That is, by comparing the primer information associated with the base sequence to be typed with the primer information of the reference sequence, the above-mentioned second candidate type combination to be analyzed is obtained.

Further, the analysis module 520 is further configured to: acquire the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed, and determine the final type of the base sequence to be typed according to the intersection. Wherein, the allelic type information of the reference sequence includes IMGT HLA type I and type II site gene information, and the site name of the base sequence to be typed included in the file information is the site gene information of the corresponding site. That is: the final type of the base sequence to be typed is confirmed according to the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed.

The display module 530 is used to display the information stored by the storage module and the analysis result of the analysis module.

In summary, the system of the embodiment of the present invention includes: a storage module (i.e. a database module), an analysis module and a display module (i.e. an interface display module), wherein all known HLA type sequences in the IMGT/HLA database will be displayed as designed The good format is stored in the static database of the storage module. At the same time, the valid data of the sample to be analyzed (base sequence to be analyzed) imported into the software will also be stored in the dynamic database of the storage module in a fixed format, which is convenient for data retrieval during analysis.

The analysis module realizes the typing function, for example: the analysis module realizes the core function of typing the HLA genotype of the sample, and there are two sub-modules: the analysis file extraction information module and the analysis sample HLA typing module. The main function of the analysis file extraction information module is to extract the useful information of the sequencing sequence peak map file and store it in the storage module. The HLA typing module of the analysis sample merges the file information and realizes the function of HLA typing according to the sample.

The display module (that is, the interface display module) generates an operation interface, that is, a visual interface. For example, the interface display module is shown in Figure 4. The sample list realizes the display of the opened files according to the structure of the file tree, and realizes the related right-click function; the type list realizes the display of the typing results, and realizes the related right-click function; base Base navigation realizes the function of quickly navigating different regions of exons; sequence display realizes the display function of aligned sequences. Electropherogram display realizes the display function of electropherogram.

According to the polymerase chain reaction product sequencing and typing system of the embodiment of the present invention, according to the reference sequence and its related information, the base sequence to be typed and its related information can directly identify the combination of the base sequence to be typed and the reference sequence. Therefore, the step of aligning the read base sequence to the corresponding position of each reference sequence is reduced, and the comparison time is shortened. Especially when screening and searching a large range of candidate types (such as many reference sequences, many types), the comparison time is greatly shortened. In addition, the re-identification and typing are carried out according to the primer information, thus ensuring the correctness and reliability of the typing. In addition, the system can automatically type the base sequence to be typed, reducing the tediousness of manually importing GSSP information, effectively saving time and reducing labor costs.

In the description of this specification, descriptions referring to the terms "one embodiment", "some embodiments", "example", "specific examples", or "some examples" mean that specific features described in connection with the embodiment or example , structure, material or characteristic is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

Although the embodiments of the present invention have been shown and described, those skilled in the art can understand that various changes, modifications and substitutions can be made to these embodiments without departing from the principle and spirit of the present invention. and modifications, the scope of the invention is defined by the appended claims and their equivalents.

Claims (12)

1. A method for polymerase chain reaction product sequencing typing, is characterized in that, comprises the following steps: Obtaining the base sequence to be typed, the file information associated with the base sequence to be typed, and the primer file information, wherein the file information includes the site name of the base sequence to be typed; identifying the alignment positional relationship between the base sequence to be typed and the reference sequence according to the file information and the allelic type information of the reference sequence, so as to obtain the first candidate type combination to be analyzed; Further typing and identifying the base sequence to be typed according to the primer file information and the primer information of the reference sequence to obtain a second candidate type combination to be analyzed; and Obtaining the final type of the base sequence to be typed according to the first combination of candidate types to be analyzed and the second combination of candidate types to be analyzed. 2. the method for polymerase chain reaction product sequencing typing according to claim 1, is characterized in that, described base to be typed is identified according to the allele type information of described file information and reference sequence The alignment positional relationship between the sequence and the reference sequence to obtain the first combination of candidate types to be analyzed, further including: matching the site name of the base sequence to be typed according to the allelic type information of the reference sequence included in the file information; Obtaining the alignment positional relationship between the base sequence to be typed and the reference sequence according to the matching result; The first combination of candidate types to be analyzed is obtained according to the alignment positional relationship. 3. the method for polymerase chain reaction product sequencing typing according to claim 2, is characterized in that, the allelic type information of described reference sequence comprises the HLA type I of IMGT and type II site gene information, The site name of the base sequence to be typed included in the file information is the site gene information of the corresponding site. 4. the method for polymerase chain reaction product sequencing typing according to claim 1, is characterized in that, described base sequence to be typed according to the primer information of described primer file information and described reference sequence Carry out further typing identification to obtain the second combination of candidate types to be analyzed, further including: matching the primer file information with the primer information of the reference sequence; If the matching is consistent, the corresponding reference sequence is added to the second combination of candidate types to be analyzed. 5. The method for sequencing and typing polymerase chain reaction products according to claim 4, wherein the primer information of the reference sequence includes GSSP primer information of HLA type I and type II, that is, spare primer information. 6. The method for sequencing and typing of polymerase chain reaction products according to any one of claims 1-5, characterized in that, according to the combination of the first candidate type to be analyzed and the second to be analyzed The combination of candidate types to obtain the final type of the base sequence to be typed further includes: Obtaining the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed; The final type of the base sequence to be typed is determined according to the intersection. 7. A system for sequencing and typing polymerase chain reaction products, characterized in that it comprises: The storage module is used to store the base sequence to be typed, the file information associated with the base sequence to be typed and the primer file information, as well as the reference sequence, the allelic type information and primer information of the reference sequence, Wherein, the file information includes the site name of the base sequence to be typed; The analysis module is used to identify the alignment position relationship between the base sequence to be typed and the reference sequence according to the file information and the allelic type information of the reference sequence, so as to obtain the first candidate type combination to be analyzed, and According to the primer file information and the primer information of the reference sequence, the base sequence to be typed is further typed and identified to obtain the second candidate type combination to be analyzed, and according to the first candidate type to be analyzed Genre combination and the second candidate type to be analyzed are combined to obtain the final type of the base sequence to be typed; and The display module is used to display the information stored by the storage module and the analysis result of the analysis module. 8. The system of polymerase chain reaction product sequencing typing according to claim 7, is characterized in that, described analysis module is used for: matching the site name of the base sequence to be typed according to the allelic type information of the reference sequence included in the file information; Obtaining the alignment positional relationship between the base sequence to be typed and the reference sequence according to the matching result; The first combination of candidate types to be analyzed is obtained according to the alignment positional relationship. 9. The system of polymerase chain reaction product sequencing typing according to claim 8, is characterized in that, the allelic type information of described reference sequence comprises the HLA type I of IMGT and type II site gene information, The site name of the base sequence to be typed included in the file information is the site gene information of the corresponding site. 10. The system of polymerase chain reaction product sequencing typing according to claim 7, is characterized in that, described analysis module is also used for: matching the primer file information with the primer information of the reference sequence; If the matching is consistent, the corresponding reference sequence is added to the second combination of candidate types to be analyzed. 11. The system for sequencing and typing polymerase chain reaction products according to claim 10, wherein the primer information of the reference sequence includes GSSP primer information of HLA type I and type II, that is, spare primer information. 12. The system for sequencing and typing polymerase chain reaction products according to any one of claims 7-11, wherein the analysis module is also used for: Obtaining the intersection of the first candidate type combination to be analyzed and the second candidate type combination to be analyzed; The final type of the base sequence to be typed is determined according to the intersection.