The very sensitive Jerne method of detecting the plaque-forming cells was used in the study of th... more The very sensitive Jerne method of detecting the plaque-forming cells was used in the study of the immunosuppressive effect of ionizing radiation on laboratory mice. A significant immunosuppression was obtained after irradiation of the individual groups of mice with 3, 4, 5 and 6 Gy. Successive regeneration of the immunity system occurred after 42 hours only in the groups of mice irradiated with 3 and 4 Gy. mice; ionizing radiation; immunoproductive cells; Jerne method.
The causative agents of respiratory diseases of turkeys represent, primarily in fattening farms, ... more The causative agents of respiratory diseases of turkeys represent, primarily in fattening farms, a substantial risk of economic and breeding problems. The purpose of this communication is to provide informa - tion on the prevalence of respiratory agents of turkeys and chickens in several fattening and production farms in Southern Moravia. This study was focused on pathogens causing bacterial diseases such as Ornithobacteriosis and Mycoplasmosis, as well as viral rhinotracheitis and laryngotracheitis of poultry. The laboratory diagnosis of these diseases has been performed in our institute since January 2008. We examined 249 samples of turkeys and chickens from a single rearing house and six fattening farms in Southern Moravia. The samples were examined using the PCR or RT-PCR method. The typing of isolates of Ornithobacterium rhinotracheale was done using the M13 fingerprinting method. We established the prevalence of pathogens such as Ornithobacterium rhinotrache- ale (ORT), Mycopl...
Adenoviruses are a widespread cause of diverse human infections with recently confirmed zoonotic ... more Adenoviruses are a widespread cause of diverse human infections with recently confirmed zoonotic roots in African great apes. We focused on savanna-dwelling chimpanzees in the Issa Valley (Tanzania), which differ from those from forested sites in many aspects of behavior and ecology. PCR targeting the DNA polymerase gene detected AdV in 36.7% (69/188) of fecal samples. We detected five groups of strains belonging to the species Human mastadenovirus E and two distinct groups within the species Human mastadenovirus C based on partial hexon sequence. All detected AdVs from the Issa Valley are related to those from nearby Mahale and Gombe National Parks, suggesting chimpanzee movements and pathogen transmission.
The intensity and distribution of typical lesions in organs and tissues were determined in 38 she... more The intensity and distribution of typical lesions in organs and tissues were determined in 38 sheep with serological positivity to the Maedi-Visna disease. The lesions in the lungs, mammary gland, CNS and joints were evaluated according to the number of marks. The lungs, mammary gland and CNS were affected most frequently. Concurrent incidence of pulmonary adenomatosis was determined in 57% of the examined cases. Differential diagnostics of typical M-V virus induced lesions and pulmonary adenomatosis is discussed in this paper. Identification of eight cases of meningoencephalitis, which is considered as the initial stage of Visna form, is a priority observation in the Czech Republic. With respect to distribution of lentivirus carriers, the authors emphasize the need of detailed pathomorphological examination of culled sheep.
In the present paper the results are presented of pathomorphological examination of 38 sacrificed... more In the present paper the results are presented of pathomorphological examination of 38 sacrificed sheep with positive reactions to the Maedi-Visna disease. Lungs, mammary gland, joints and other organs with macroscopical lesions were subjected to histopathological examination. The CNS was examined by the method of a complete series of frontal sections. Diagnostics of M-V typical lesions revealed them in 87% of the examined sheep, in 55.2% of examined lungs, in 58.6% of examined mammary glands, 27.3% examined CNS and 4.7% examined joints. In 57.1% of the cases, M-V lesions of the lungs were complicated by simultaneous incidence of pulmonary adenomatosis, chronic purulent or granulomatous pneumonia. No clinical symptoms were found in a majority of sheep, while the M-V typical lesions of organs were not determined in about fifty percent of sheep. The observed morphological lesions corresponded to initial or medium progressive affection. A proof of precipitation antibodies and identific...
A group of five ferrets vaccinated against the canine distemper virus (CDV) was evaluated as to t... more A group of five ferrets vaccinated against the canine distemper virus (CDV) was evaluated as to the onset of anti-CDV antibody production and the serum levels of the animals were monitored for one year. The ferrets were immunized with a live attenuated vaccine. The vaccination pattern was as follows: primary vaccination at the age of 6 weeks, fi rst revaccination at 30 days after primary vaccination, and second revaccination after another 30 days. Blood samples were taken prior to primary vaccination and then at 30-day intervals (sampling 1 to 12). The whole experimental cycle covered the period of one year from primary vaccination (till the age of 1 year and 6 weeks). Serum samples were analysed for anti-CDV virus-neutralisation antibodies using a virus-neutralisation test using the Onderstepoort CDV strain. All ferrets had zero virus-neutralisation antibody titres before primary vaccination. Two ferrets produced virus-neutralisation antibodies as a response to first revaccination....
This study reports on the first quantification of avian influenza virus in the organs of mute swa... more This study reports on the first quantification of avian influenza virus in the organs of mute swans that died during the epizootic of avian influenza (H5N1) between January and April 2006 in the Czech Republic. The quantitative real-time Reverse Transcriptase PCR (qRT-PCR) assay based on a TaqMan probe was developed for a rapid detection and quantification of avian influenza virus RNA in clinical samples collected from mute swans. Conserved regions in the matrix protein gene of avian influenza virus served as targets for the primers and TaqMan probe design. A recombinant plasmid containing the matrix protein gene amplicon was constructed for a quantitative assay of copy numbers of the target gene. Quantification of avian influenza virus RNA was accomplished using a standard curve generated from ten-fold serial dilutions of recombinant plasmid DNA in the range of 102 to 108 copies/µl. Avian influenza virus A/Cygnus olor/Brno-cz/2006 was adapted to grow in VERO cells. In the same pass...
Porcine reproductive and respiratory syndrome (PRRS) virus represents a major threat to the swine... more Porcine reproductive and respiratory syndrome (PRRS) virus represents a major threat to the swine industry worldwide. This study describes the transmission of a European strain of PRRS-1 to a pig farm leading to the spread of the virus to different age categories of pigs and the development of clinical signs in pregnant sows and piglets. Porcine reproductive and respiratory syndrome aetiology was confirmed by serological tests and by virus isolation and subsequent sequencing. Repeated mass vaccination using modified live virus was used to synchronise the level of PRRS-specific immunity in all age categories of animals and to hinder virus circulation in the herd. Four months after the second mass vaccination, regular modified live virus vaccination of sows and gilts was implemented. Introduction of sentinel animals demonstrated cessation of virus circulation and the success of the control programme.
Between 2001 and 2003, a total of 194 samples of brain tissues of wild mustelids from the Czech R... more Between 2001 and 2003, a total of 194 samples of brain tissues of wild mustelids from the Czech Republic were tested for the presence of canine distemper virus (CDV) by direct immunofluorescence examination. Out of 21 animals exhibiting symptoms of the disease or changed behaviour, one mustelid was CDV positive (5% prevalence). In this group, 1 out of 18 stone martens (Martes foina) was CDV positive, while 2 pine martens (Martes martes) and 1 Eurasian badger (Meles meles) were CDV negative. Of 173 animals with unknown case history, 1 sample was positive (0.6% prevalence). In this group of animals, 1 out of 19 Eurasian badgers was positive, and stone martens (n = 96), pine martens (n = 4), polecats (Mustela putorius) ((n = 28), steppe polecats (Mustela eversmani) (n = 4), common weasels (Mustela nivalis) (n = 4), stoats (Mustela erminea) (n = 3) and American minks (Mustela vison) (n = 19) were negative. Clinical distemper was demonstrated in three stone marten pup siblings. In two of...
The most adequate means of diagnosing infections with caprine arthritis encephalitis (CAE) and Ma... more The most adequate means of diagnosing infections with caprine arthritis encephalitis (CAE) and Maedi-Visna (MV) viruses is the demonstration of antibodies to these viruses in milk or in blood serum. Various techniques and a range of different antigen preparations can be used for this purpose. We have evaluated two different whole virus antigen preparations derived from lamb synovial cells infected with CAE and MV viruses and recombinant antigen containing the capsid protein expressed in E. coli in ELISA test. The suitability of different detergents for solubilization of whole virus particles is shown in Tab. I. The detergents used were ether (50%, 10 min, 37 degrees C), octyl-beta-D glucopyranoside - OGP (2%, 2 h, 37 degrees C), and sodium dodecylsulphate SDS (0.125%, 10 min, 37 degrees C). Antigens prepared with SDS gave the best results and were then used in the following antigen comparative study. All antigens (whole virus and recombinant core protein) were used for the coating o...
In the present paper antigen preparation is described to be used for diagnostics of Maedi-Visna d... more In the present paper antigen preparation is described to be used for diagnostics of Maedi-Visna disease by immunodiffusion test (IDT). The antigen was prepared by cultivation of the Maedi-Visna virus, OLV strain, in cell culture of the synovial membrane of carpal joint and plexus choroides of lamb. Cell cultures were used for 10-25 subcultures. The antigen was concentrated by polyethylene glycol application. The characteristics of the prepared diagnostic antigen were evaluated by its comparison with a reference, commercial preparation. Sufficient sensitivity and specificity of the prepared antigen was demonstrated by testing in a sample of 100 ovine serums.
On five newly established large goat farms the incidence of antibodies to some chronic and latent... more On five newly established large goat farms the incidence of antibodies to some chronic and latent infections (arthritis and encephalitis, Q-fever, caseous lymphadenitis and toxoplazmosis) was investigated in the first six months of the year 1994. Agar-gel immunodiffusion did not reveal any antibodies to arthritis and encephalitis of goats (CAE). Complement fixation test did not demonstrate any antibodies to Q-fever. Neither agar-gel immunodiffusion nor neutralization test confirmed any antibodies to caseous lymphadentitis. Complement fixation test (titer 1:8 and more) revealed antibodies to toxoplazmosis at 20.2% of the cases. No clinical symptoms of toxoplazmosis were observed on the investigated goat farms. It was recommended to take a preventive serological examination of goats against CAE, Q-fever and caseous lymphadenitis before they were housed on the given farms. By maintaining high zoohygienic parameters on the large goat farms it is possible to except a decrease in prevalen...
The aim of this work was to express recombinant nonstructural Nsp7 protein of European genotype o... more The aim of this work was to express recombinant nonstructural Nsp7 protein of European genotype of porcine reproductive and respiratory syndrome virus and to evaluate its diagnostic sensitivity and specificity in serological diagnostics of the disease. The gene coding for Nsp7 protein was expressed in Escherichia coli cells and purified by IMAC. Serological reactivity of purified protein was assessed on a panel of swine sera in indirect ELISA test. Serum samples originated from PRRS positive farms, herds free of PRRS infection and PRRS free herds vaccinated with an inactivated vaccine. Nsp7 antigen proved to be suitable for serological detection of PRRS specific antibodies, showing diagnostic sensitivity of 82.2% and specificity of 97.6% when compared with IDEXX ELISA test. The nonstructural protein proved to be suitable for use as an antigen for the differentiation of post-infection and post-vaccination antibodies in pigs vaccinated with the inactivated vaccine. But the low overall antibody response to N protein after this type of vaccination makes this concept rather theoretical.
ABSTRACT A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lent... more ABSTRACT A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lentivirus (OvLV) in peripheral blood mononuclear cells was developed. Primers for snPCR were situated within the gag gene of the Maedi-Visna virus (MVV) genome. A comparison between the snPCR and serological tests (agar gel immunodiffusion test, immunoblot) were performed using 98 ovine blood samples. Thirty (30.6%) of the 98 sheep examined had antibodies specific for the MVV. PCR showed 21 of them to be positive and nine seropositive animals to be PCR negative. Six of the 68 serologically negative sheep were found to be PCR positive, probably due to delayed seroconversion. The PCR amplification products of these six sheep were sequenced and subjected to phylogenetic analysis. The resulting phylogenetic tree of partial gag gene sequences confirmed that the ovine lentivirus genotype in the Czech Republic is more closely related to the prototype MVV isolates than to the caprine arthritis encephalitis viruses.
Three oligonucleotide primers for semi-nested polymerase chain reaction (PCR) were designed accor... more Three oligonucleotide primers for semi-nested polymerase chain reaction (PCR) were designed according to already published sequences of porcine circovirus types 1 (PCV-1) and 2 (PCV-2) isolates. These primers were used to detect PCV-2 DNA. A positive amplification reaction was visualized from a DNA suspension containing as few as 10 copies of virus DNA. In total. 77 samples of inguinal lymph nodes and nasal swabs from pigs in the Czech Republic were used to detect the virus. Thirty-seven of them were positive for PCV-2 DNA. In order to confirm specificity of the PCR reaction, seven DNA fragments were sequenced. Czech PCV sequences were found to have a 92-97% homology with other known PCV-2 strains and only 80-83% homology with PCV-1 strains.
The very sensitive Jerne method of detecting the plaque-forming cells was used in the study of th... more The very sensitive Jerne method of detecting the plaque-forming cells was used in the study of the immunosuppressive effect of ionizing radiation on laboratory mice. A significant immunosuppression was obtained after irradiation of the individual groups of mice with 3, 4, 5 and 6 Gy. Successive regeneration of the immunity system occurred after 42 hours only in the groups of mice irradiated with 3 and 4 Gy. mice; ionizing radiation; immunoproductive cells; Jerne method.
The causative agents of respiratory diseases of turkeys represent, primarily in fattening farms, ... more The causative agents of respiratory diseases of turkeys represent, primarily in fattening farms, a substantial risk of economic and breeding problems. The purpose of this communication is to provide informa - tion on the prevalence of respiratory agents of turkeys and chickens in several fattening and production farms in Southern Moravia. This study was focused on pathogens causing bacterial diseases such as Ornithobacteriosis and Mycoplasmosis, as well as viral rhinotracheitis and laryngotracheitis of poultry. The laboratory diagnosis of these diseases has been performed in our institute since January 2008. We examined 249 samples of turkeys and chickens from a single rearing house and six fattening farms in Southern Moravia. The samples were examined using the PCR or RT-PCR method. The typing of isolates of Ornithobacterium rhinotracheale was done using the M13 fingerprinting method. We established the prevalence of pathogens such as Ornithobacterium rhinotrache- ale (ORT), Mycopl...
Adenoviruses are a widespread cause of diverse human infections with recently confirmed zoonotic ... more Adenoviruses are a widespread cause of diverse human infections with recently confirmed zoonotic roots in African great apes. We focused on savanna-dwelling chimpanzees in the Issa Valley (Tanzania), which differ from those from forested sites in many aspects of behavior and ecology. PCR targeting the DNA polymerase gene detected AdV in 36.7% (69/188) of fecal samples. We detected five groups of strains belonging to the species Human mastadenovirus E and two distinct groups within the species Human mastadenovirus C based on partial hexon sequence. All detected AdVs from the Issa Valley are related to those from nearby Mahale and Gombe National Parks, suggesting chimpanzee movements and pathogen transmission.
The intensity and distribution of typical lesions in organs and tissues were determined in 38 she... more The intensity and distribution of typical lesions in organs and tissues were determined in 38 sheep with serological positivity to the Maedi-Visna disease. The lesions in the lungs, mammary gland, CNS and joints were evaluated according to the number of marks. The lungs, mammary gland and CNS were affected most frequently. Concurrent incidence of pulmonary adenomatosis was determined in 57% of the examined cases. Differential diagnostics of typical M-V virus induced lesions and pulmonary adenomatosis is discussed in this paper. Identification of eight cases of meningoencephalitis, which is considered as the initial stage of Visna form, is a priority observation in the Czech Republic. With respect to distribution of lentivirus carriers, the authors emphasize the need of detailed pathomorphological examination of culled sheep.
In the present paper the results are presented of pathomorphological examination of 38 sacrificed... more In the present paper the results are presented of pathomorphological examination of 38 sacrificed sheep with positive reactions to the Maedi-Visna disease. Lungs, mammary gland, joints and other organs with macroscopical lesions were subjected to histopathological examination. The CNS was examined by the method of a complete series of frontal sections. Diagnostics of M-V typical lesions revealed them in 87% of the examined sheep, in 55.2% of examined lungs, in 58.6% of examined mammary glands, 27.3% examined CNS and 4.7% examined joints. In 57.1% of the cases, M-V lesions of the lungs were complicated by simultaneous incidence of pulmonary adenomatosis, chronic purulent or granulomatous pneumonia. No clinical symptoms were found in a majority of sheep, while the M-V typical lesions of organs were not determined in about fifty percent of sheep. The observed morphological lesions corresponded to initial or medium progressive affection. A proof of precipitation antibodies and identific...
A group of five ferrets vaccinated against the canine distemper virus (CDV) was evaluated as to t... more A group of five ferrets vaccinated against the canine distemper virus (CDV) was evaluated as to the onset of anti-CDV antibody production and the serum levels of the animals were monitored for one year. The ferrets were immunized with a live attenuated vaccine. The vaccination pattern was as follows: primary vaccination at the age of 6 weeks, fi rst revaccination at 30 days after primary vaccination, and second revaccination after another 30 days. Blood samples were taken prior to primary vaccination and then at 30-day intervals (sampling 1 to 12). The whole experimental cycle covered the period of one year from primary vaccination (till the age of 1 year and 6 weeks). Serum samples were analysed for anti-CDV virus-neutralisation antibodies using a virus-neutralisation test using the Onderstepoort CDV strain. All ferrets had zero virus-neutralisation antibody titres before primary vaccination. Two ferrets produced virus-neutralisation antibodies as a response to first revaccination....
This study reports on the first quantification of avian influenza virus in the organs of mute swa... more This study reports on the first quantification of avian influenza virus in the organs of mute swans that died during the epizootic of avian influenza (H5N1) between January and April 2006 in the Czech Republic. The quantitative real-time Reverse Transcriptase PCR (qRT-PCR) assay based on a TaqMan probe was developed for a rapid detection and quantification of avian influenza virus RNA in clinical samples collected from mute swans. Conserved regions in the matrix protein gene of avian influenza virus served as targets for the primers and TaqMan probe design. A recombinant plasmid containing the matrix protein gene amplicon was constructed for a quantitative assay of copy numbers of the target gene. Quantification of avian influenza virus RNA was accomplished using a standard curve generated from ten-fold serial dilutions of recombinant plasmid DNA in the range of 102 to 108 copies/µl. Avian influenza virus A/Cygnus olor/Brno-cz/2006 was adapted to grow in VERO cells. In the same pass...
Porcine reproductive and respiratory syndrome (PRRS) virus represents a major threat to the swine... more Porcine reproductive and respiratory syndrome (PRRS) virus represents a major threat to the swine industry worldwide. This study describes the transmission of a European strain of PRRS-1 to a pig farm leading to the spread of the virus to different age categories of pigs and the development of clinical signs in pregnant sows and piglets. Porcine reproductive and respiratory syndrome aetiology was confirmed by serological tests and by virus isolation and subsequent sequencing. Repeated mass vaccination using modified live virus was used to synchronise the level of PRRS-specific immunity in all age categories of animals and to hinder virus circulation in the herd. Four months after the second mass vaccination, regular modified live virus vaccination of sows and gilts was implemented. Introduction of sentinel animals demonstrated cessation of virus circulation and the success of the control programme.
Between 2001 and 2003, a total of 194 samples of brain tissues of wild mustelids from the Czech R... more Between 2001 and 2003, a total of 194 samples of brain tissues of wild mustelids from the Czech Republic were tested for the presence of canine distemper virus (CDV) by direct immunofluorescence examination. Out of 21 animals exhibiting symptoms of the disease or changed behaviour, one mustelid was CDV positive (5% prevalence). In this group, 1 out of 18 stone martens (Martes foina) was CDV positive, while 2 pine martens (Martes martes) and 1 Eurasian badger (Meles meles) were CDV negative. Of 173 animals with unknown case history, 1 sample was positive (0.6% prevalence). In this group of animals, 1 out of 19 Eurasian badgers was positive, and stone martens (n = 96), pine martens (n = 4), polecats (Mustela putorius) ((n = 28), steppe polecats (Mustela eversmani) (n = 4), common weasels (Mustela nivalis) (n = 4), stoats (Mustela erminea) (n = 3) and American minks (Mustela vison) (n = 19) were negative. Clinical distemper was demonstrated in three stone marten pup siblings. In two of...
The most adequate means of diagnosing infections with caprine arthritis encephalitis (CAE) and Ma... more The most adequate means of diagnosing infections with caprine arthritis encephalitis (CAE) and Maedi-Visna (MV) viruses is the demonstration of antibodies to these viruses in milk or in blood serum. Various techniques and a range of different antigen preparations can be used for this purpose. We have evaluated two different whole virus antigen preparations derived from lamb synovial cells infected with CAE and MV viruses and recombinant antigen containing the capsid protein expressed in E. coli in ELISA test. The suitability of different detergents for solubilization of whole virus particles is shown in Tab. I. The detergents used were ether (50%, 10 min, 37 degrees C), octyl-beta-D glucopyranoside - OGP (2%, 2 h, 37 degrees C), and sodium dodecylsulphate SDS (0.125%, 10 min, 37 degrees C). Antigens prepared with SDS gave the best results and were then used in the following antigen comparative study. All antigens (whole virus and recombinant core protein) were used for the coating o...
In the present paper antigen preparation is described to be used for diagnostics of Maedi-Visna d... more In the present paper antigen preparation is described to be used for diagnostics of Maedi-Visna disease by immunodiffusion test (IDT). The antigen was prepared by cultivation of the Maedi-Visna virus, OLV strain, in cell culture of the synovial membrane of carpal joint and plexus choroides of lamb. Cell cultures were used for 10-25 subcultures. The antigen was concentrated by polyethylene glycol application. The characteristics of the prepared diagnostic antigen were evaluated by its comparison with a reference, commercial preparation. Sufficient sensitivity and specificity of the prepared antigen was demonstrated by testing in a sample of 100 ovine serums.
On five newly established large goat farms the incidence of antibodies to some chronic and latent... more On five newly established large goat farms the incidence of antibodies to some chronic and latent infections (arthritis and encephalitis, Q-fever, caseous lymphadenitis and toxoplazmosis) was investigated in the first six months of the year 1994. Agar-gel immunodiffusion did not reveal any antibodies to arthritis and encephalitis of goats (CAE). Complement fixation test did not demonstrate any antibodies to Q-fever. Neither agar-gel immunodiffusion nor neutralization test confirmed any antibodies to caseous lymphadentitis. Complement fixation test (titer 1:8 and more) revealed antibodies to toxoplazmosis at 20.2% of the cases. No clinical symptoms of toxoplazmosis were observed on the investigated goat farms. It was recommended to take a preventive serological examination of goats against CAE, Q-fever and caseous lymphadenitis before they were housed on the given farms. By maintaining high zoohygienic parameters on the large goat farms it is possible to except a decrease in prevalen...
The aim of this work was to express recombinant nonstructural Nsp7 protein of European genotype o... more The aim of this work was to express recombinant nonstructural Nsp7 protein of European genotype of porcine reproductive and respiratory syndrome virus and to evaluate its diagnostic sensitivity and specificity in serological diagnostics of the disease. The gene coding for Nsp7 protein was expressed in Escherichia coli cells and purified by IMAC. Serological reactivity of purified protein was assessed on a panel of swine sera in indirect ELISA test. Serum samples originated from PRRS positive farms, herds free of PRRS infection and PRRS free herds vaccinated with an inactivated vaccine. Nsp7 antigen proved to be suitable for serological detection of PRRS specific antibodies, showing diagnostic sensitivity of 82.2% and specificity of 97.6% when compared with IDEXX ELISA test. The nonstructural protein proved to be suitable for use as an antigen for the differentiation of post-infection and post-vaccination antibodies in pigs vaccinated with the inactivated vaccine. But the low overall antibody response to N protein after this type of vaccination makes this concept rather theoretical.
ABSTRACT A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lent... more ABSTRACT A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lentivirus (OvLV) in peripheral blood mononuclear cells was developed. Primers for snPCR were situated within the gag gene of the Maedi-Visna virus (MVV) genome. A comparison between the snPCR and serological tests (agar gel immunodiffusion test, immunoblot) were performed using 98 ovine blood samples. Thirty (30.6%) of the 98 sheep examined had antibodies specific for the MVV. PCR showed 21 of them to be positive and nine seropositive animals to be PCR negative. Six of the 68 serologically negative sheep were found to be PCR positive, probably due to delayed seroconversion. The PCR amplification products of these six sheep were sequenced and subjected to phylogenetic analysis. The resulting phylogenetic tree of partial gag gene sequences confirmed that the ovine lentivirus genotype in the Czech Republic is more closely related to the prototype MVV isolates than to the caprine arthritis encephalitis viruses.
Three oligonucleotide primers for semi-nested polymerase chain reaction (PCR) were designed accor... more Three oligonucleotide primers for semi-nested polymerase chain reaction (PCR) were designed according to already published sequences of porcine circovirus types 1 (PCV-1) and 2 (PCV-2) isolates. These primers were used to detect PCV-2 DNA. A positive amplification reaction was visualized from a DNA suspension containing as few as 10 copies of virus DNA. In total. 77 samples of inguinal lymph nodes and nasal swabs from pigs in the Czech Republic were used to detect the virus. Thirty-seven of them were positive for PCV-2 DNA. In order to confirm specificity of the PCR reaction, seven DNA fragments were sequenced. Czech PCV sequences were found to have a 92-97% homology with other known PCV-2 strains and only 80-83% homology with PCV-1 strains.
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