TWI818398B - Convective polymerase chain reaction apparatus and optical detecting method thereof - Google Patents

Abstract

A convective polymerase chain reaction apparatus and an optical detecting method thereof are provided. The optical detecting method includes the following steps. The substance in the tube to be tested is heated. At least two monochromatic lights are provided and combined by a light combining element and then to irradiate the tube to be tested. At least two kinds of excited lights generated by the excitation of the substance in the tube to be tested by the at least two monochromatic lights are sensed.

Description

Thermal convection polymerase chain reaction device and its optical detection method

The present disclosure relates to a reaction device and a detection method, and in particular to a thermal convection polymerase chain reaction device and an optical detection method thereof.

With the prevalence of various infectious diseases, rapid and accurate virus detection methods are urgently needed to prevent and monitor infectious disease pandemics. Viral nucleic acid detection using nucleic acid extraction and polymerase chain reaction can detect whether the subject carries viral gene fragments in the body. This detection method is highly sensitive and can detect only a small amount of virus in the subject's body.

In the thermal convection polymerase chain reaction, the thermal convection principle is used to directly heat the reaction vessel containing the liquid to be measured, causing a temperature gradient in the liquid to be measured and causing thermal convection. By causing the liquid to be tested to continuously change its temperature during the thermal convection process, the thermal convection of the liquid to be tested can amplify nucleic acids, shorten the time required for temperature changes in traditional methods, and has the advantages of simple structure and low cost of the testing device. . The thermal convection polymerase chain reaction device currently used is made of capillary capillaries made of plastic or glass. The test tube serves as a reaction vessel, and the test tube is irradiated with light to excite the fluorescent reagent in the tube to generate a fluorescent signal, and then the fluorescent signal is sensed by the sensor to achieve the purpose of monitoring the polymerase chain reaction.

Polymerase chain reaction is a molecular biology technique used to amplify specific deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) fragments. A typical polymerase chain reaction device uses a thermocycler to repeatedly heat and cool to repeatedly perform thermal cycle steps. The thermal energy is gradually conducted (thermal conduction) through the metal block into the reaction tube, so that the DNA or ribonucleic acid fragments can denature (95°C), adhere (45°C-65°C) and extend (72°C) at different temperatures. ) and three different temperature cycles. However, when there is more than one type of target virus to be detected, each device can only detect one type of virus in one tube to be tested. The same number of tubes to be tested needs to be collected according to the number of types of target viruses to increase the number of test tubes to be tested. the burden of the person.

The present disclosure provides a thermal convection polymerase chain reaction device and an optical detection method thereof, which can detect multiple targets at the same time, thereby reducing the overall required amount of specimens.

The thermal convection polymerase chain reaction device of the present disclosure includes a base, a heating element, a light combining element, at least two light-emitting elements and at least two light sensors. The number of light sensors corresponds to the number of light-emitting elements. The base body is suitable for carrying a tube to be tested. The heating element is installed on the base to heat the tube to be tested through the base. The light combining element includes a connected first plate-shaped part, a second plate-shaped part, a third plate-shaped part and a first plate-shaped part. Four plates. The first plate-shaped part and the second plate-shaped part define a light emitting area. The second plate-shaped part and the third plate-shaped part define a first light incident area. The third plate-shaped part and the fourth plate-shaped part define a second light incident area. The first plate-shaped part and the fourth plate-shaped part define a third light incident area. The light-emitting element is at least two of a first light-emitting element, a second light-emitting element and a third light-emitting element. The first light-emitting element is adapted to provide a first color light toward the first light incident area. The second light-emitting element is adapted to provide a second color light toward the second light incident area. The third light-emitting element is adapted to provide a third color light toward the third light incident area. The first plate-shaped portion is adapted to reflect the first color light and allow the second color light and the third color light to pass. The second plate-shaped portion is adapted to reflect the third color light and allow the first color light and the second color light to pass. The third plate-shaped portion is adapted to reflect the first color light and allow the second color light to pass through. The fourth plate-shaped portion is adapted to reflect the third color light and allow the second color light to pass through. At least two of the first color light, the second color light and the third color light enter the light emitting area and emit light toward the base to illuminate the tube to be tested. At least two of the first photo sensor, the second photo sensor and the third photo sensor are installed on the base. The light sensor is installed on the base and is at least two of a first light sensor, a second light sensor and a third light sensor. The first light sensor is adapted to receive a first excitation light generated when the substance in the tube to be tested is excited by the first color light. The second light sensor is adapted to receive a second excitation light generated when the substance in the tube to be tested is excited by the second color light. The third light sensor is adapted to receive a third excitation light generated when the substance in the tube to be tested is excited by the third color light.

The optical detection method of the present disclosure uses the above-mentioned thermal convection polymerase chain reaction device for detection. The method includes the following steps of heating the substance in the tube to be tested. Provide at least two of the first color light, the second color light and the third color light and use combined light The components are combined with light, and then the tube to be tested is irradiated. Sensing at least one of the first excitation light generated by the substance in the tube to be tested when excited by the first color light, the second excitation light generated by the second color light, and the third excitation light generated by the third color light. Two.

Based on the above, in the thermal convection polymerase chain reaction device and its optical detection method disclosed in the present disclosure, at least two colors of light can be detected simultaneously, thereby reducing the amount of specimen used.

50: Tube to be tested

60:Spectrometer

100: Thermal convection polymerase chain reaction device

110: base body

112: Slender through hole

130:Heating element

140: Light combining element

140A: First plate-shaped part

140B: Second plate-shaped part

140C: The third plate-shaped part

140D: The fourth plate-like part

140E: Fixed seat

150B: First light-emitting element

150G: Second light-emitting element

150R: The third light-emitting element

150B1,150G1,150R1: LED

150B2,150G2,150R2: Optical filter

160B: First light sensor

160G: Second light sensor

160R: Third light sensor

162B: First filter element

162G: Second filter element

162R: The third filter element

170:Circuit board

180: Condensing unit

182: Lens tube

184: condenser lens

A10,A20: long axis

R12: light emission area

R14: first light entrance area

R16: Second light incident area

R18: The third light entrance area

LB: first color light

LG: Second color light

LR: third color light

EB: first excitation light

EG: second excitation light

ER: third excitation light

Figure 1 is a three-dimensional cross-sectional view of a thermal convection polymerase chain reaction device according to an embodiment of the present disclosure.

FIG. 2 is a partial top view of the thermal convection polymerase chain reaction device of FIG. 1 .

FIG. 3 is a schematic diagram illustrating the function of the light condensing unit of the thermal convection polymerase chain reaction device of FIG. 1 .

Figure 4 is a schematic diagram of the light condensing unit of the thermoconvection polymerase chain reaction device of Figure 3 after adjustment.

FIG. 5 is an exploded schematic diagram of the light combining element of the thermoconvection polymerase chain reaction device of FIG. 1 .

FIG. 6 is a partial top view of the light combining element of FIG. 5 .

Figure 1 is a three-dimensional cross-sectional view of a thermal convection polymerase chain reaction device according to an embodiment of the present disclosure. FIG. 2 is a partial top view of the thermal convection polymerase chain reaction device of FIG. 1 , and some components of the thermal convection polymerase chain reaction device are omitted in FIG. 2 . Please refer to FIGS. 1 and 2 . The thermal convection polymerase chain reaction device 100 of this embodiment is suitable for performing thermal convection polymerase chain reaction and can amplify specific DNA or ribonucleic acid fragments for virus detection. However, The thermal convection polymerase chain reaction device 100 of the present disclosure is not limited to long-term use.

The thermal convection polymerase chain reaction device 100 includes a base 110, a heating element 130, a light combining element 140, at least two light emitting elements and at least two light sensors. The number of light sensors is set corresponding to the number of light-emitting elements. As shown in FIG. 1 , the light-emitting element can be at least two of a first light-emitting element 150B, a second light-emitting element 150G and a third light-emitting element 150R; the light sensor can be a first light sensor 160B. , at least two of a second photo sensor 160G and a third photo sensor 160R. In other words, the light-emitting elements and light sensors in the thermal convection polymerase chain reaction device 100 of this embodiment may only include the first light-emitting element 150B, the second light-emitting element 150G, the first light sensor 160B and the second light sensor. The detector 160G may only include the third light-emitting element 150R, the second light-emitting element 150G, the third light sensor 160R and the second light sensor 160G, or only the first light-emitting element 150B, the third light-emitting element 150R, The first light sensor 160B and the third light sensor 160R may also include a first light emitting element 150B, a second light emitting element 150G, a third light emitting element 150R, a first light sensor 160B, a second light sensor. The detector 160G and the third photo sensor 160R. In the following, the thermal convection polymerase chain reaction device 100 will also include the first light-emitting element. The device 150B, the second light-emitting element 150G, the third light-emitting element 150R, the first light sensor 160B, the second light sensor 160G and the third light sensor 160R are taken as an example for illustration.

The base 110 is suitable for carrying a tube 50 to be tested. The heating element 130 is installed on the base 110 to heat the tube 50 to be tested through the base 110 . The light combining element 140 includes a first plate-shaped part 140A, a second plate-shaped part 140B, a third plate-shaped part 140C and a fourth plate-shaped part 140D that are connected. Specifically, each of the first plate-shaped portion 140A, the second plate-shaped portion 140B, the third plate-shaped portion 140C and the fourth plate-shaped portion 140D is connected to the other three through one surface, so as to be arranged to form an X-shaped structure. The first plate-shaped part 140A and the second plate-shaped part 140B define a light emitting area R12. The second plate-shaped portion 140B and the third plate-shaped portion 140C define a first light incident region R14. The third plate-shaped portion 140C and the fourth plate-shaped portion 140D define a second light incident region R16. The first plate-shaped portion 140A and the fourth plate-shaped portion 140D define a third light incident region R18.

The first light-emitting element 150B is suitable for providing a first color light LB toward the first light incident area R14. The second light emitting element 150G is suitable for providing a second color light LG toward the second light incident area R16. The third light-emitting element 150R is suitable for providing a third color light LR toward the third light incident area R18. The first plate-shaped portion 140A is adapted to reflect the first color light LB and allow the second color light LG and the third color light LR to pass. The second plate-shaped portion 140B is adapted to reflect the third color light LR and allow the first color light LB and the second color light LG to pass. The third plate portion 140C is adapted to reflect the first color light LB and allow the second color light LG to pass. The fourth plate portion 140D is adapted to reflect the third color light LR and allow the second color light LG to pass. Therefore, the first plate-shaped portion 140A and the third plate-shaped portion 140C can be dichroic mirrors with the same optical properties, and the second plate-shaped portion 140B and the fourth plate-shaped portion 140D can both be dichroic mirrors with another optical property. Dichroic mirror. At least two of the first color light LB, the second color light LG and the third color light LR enter from the corresponding first light incident area R14, the second light incident area R16 and the third light incident area R18. The light combining element 140 enters the light emitting area R12 through the optical effects (such as reflection and penetration) of the first plate-like part 140A, the second plate-like part 140B, the third plate-like part 140C and the fourth plate-like part 140D, And emits light toward the base 110 to illuminate the tube 50 to be tested.

Based on the X-shaped structural design of the light combining element 140, after entering the first light incident area R14, the first color light LB can directly pass through the second plate-shaped portion 140B to the first plate-shaped portion 140A, and be reflected by the first plate-shaped portion 140A. Then it enters the light emitting area R12, and may also be reflected by the third plate-shaped part 140C and then pass through the second plate-shaped part 140B to enter the light emitting area R12. After entering the second light entrance area R16, the second color light LG can directly pass through the third plate-shaped portion 140C and the second plate-shaped portion 140B and enter the light-emitting area R12, or can directly pass through the fourth plate-shaped portion 140D and the first plate. The shape portion 140A enters the light emitting area R12. After entering the third light incident area R18, the third color light LR can directly pass through the first plate-shaped portion 140A to the second plate-shaped portion 140B, be reflected by the second plate-shaped portion 140B, and then enter the light exit area R12, or can also be reflected by the fourth plate-shaped portion 140B. After reflection from the plate-shaped portion 140D, it passes through the first plate-shaped portion 140A and enters the light emitting area R12. Therefore, the first color light LB, the second color light LG and the third color light LR that enter different light entrance areas all enter the light exit area R12 and emit light toward the base 110 to illuminate the tube to be tested 50 .

At least two of the first photo sensor 160B, the second photo sensor 160G and the third photo sensor 160R are installed on the base 110 . The first light sensor 160B is adapted to receive a first excitation light EB generated when the substance in the tube to be tested 50 is excited by the first color light LB. The second light sensor 160G is adapted to receive the substance in the tube 50 to be tested. A second excitation light EG is generated by excitation of the second color light LG. The third light sensor 160R is adapted to receive a third excitation light ER generated when the substance in the tube to be tested 50 is excited by the third color light LR.

The optical detection method of this embodiment can be detected using the above-mentioned thermal convection polymerase chain reaction device 100, and the method includes the following steps. Heating the substance in the tube to be tested 50 causes the substance in the tube to be tested 50 to generate a temperature gradient and cause a thermal convection phenomenon, thereby causing nucleic acid amplification. In addition, at least two of the first color light LB, the second color light LG and the third color light LR are provided, and the light combining element 140 is used to combine the light and then illuminate the tube to be tested 50 . Then, the material in the tube to be tested 50 is sensed to be generated by the first excitation light EB excited by the first color light LB, the second excitation light EG generated by the second color light LG, and the third color light LR. The third excitation light ER is at least two of them.

In the thermal convection polymerase chain reaction device 100 of this embodiment, due to the use of the light combining element 140, multiple colored lights can be simultaneously illuminated on the tube to be tested 50, thereby allowing different substances in the tube to be tested 50 to be excited by different colored lights. Produce different excitation lights. As a result, multiple substances in the tube to be tested 50 can be detected at the same time, thereby reducing the number of inspections and reducing the burden on the subject.

In addition, the light combining element 140 of this embodiment has an X-shaped structure composed of a first plate-shaped portion 140A, a second plate-shaped portion 140B, a third plate-shaped portion 140C and a fourth plate-shaped portion 140D. Compared with solid cubic light combining elements, they are lighter in weight and can also reduce the problems caused by total reflection. In addition, because the light combining element 140 is separated from the base 110, the first light-emitting element 150B and the third light-emitting element 150B next to the light combining element 140 The second light-emitting element 150G and the third light-emitting element 150R will also be far away from the heating element 130 next to the base 110, thus preventing the first light-emitting element 150B, the second light-emitting element 150G and the third light-emitting element 150R from being heated and changing the light-emitting characteristics.

The base 110 of this embodiment is suitable for carrying the tube to be tested 50 in its center. Specifically, the tube 50 to be tested is arranged in the base 110 in the vertical and horizontal directions, and the first photo sensor 160B, the second photo sensor 160G and the third photo sensor 160R are respectively arranged in the base 110 The heating element 130 is located on the other side of the base 110 . In this embodiment, the base 110 is provided with the first light sensor 160B, the second light sensor 160G, the third light sensor 160R and the heating element 130 on four surfaces surrounding the tube to be tested 50 . In other words, the first photo sensor 160B, the second photo sensor 160G, the third photo sensor 160R and the heating element 130 are arranged around the tube to be tested 50 through the base 110 . The first photo sensor 160B, the second photo sensor 160G and the third photo sensor 160R respectively receive the first excitation light EB, the second excitation light EG and the third excitation light ER and then transmit the signals to the spectrometer 60 Analysis is performed to confirm whether there is a substance set as a detection target in the tube 50 to be tested.

In this embodiment, the thermal convection polymerase chain reaction device 100 further includes at least two filter elements. The filter element may be at least two of a first filter element 162B, a second filter element 162G, and a third filter element 162R. The number of filter elements is set corresponding to the number of photo sensors. For example, if the thermal convection polymerase chain reaction device 100 includes the first photo sensor 160B, then the first filter element 162B can be provided correspondingly, such as thermal convection polymerase chain reaction device 100. The convective polymerase chain reaction device 100 includes a second light sensor 160G, and a second filter element 162G can be provided correspondingly, such as thermal convection polymerization. The enzyme chain reaction device 100 includes a third light sensor 160R, and a third filter element 162R can be provided corresponding thereto. Specifically, the first filter element 162B is disposed between the first photo sensor 160B and the base 110 . The second filter element 162G is disposed between the second photo sensor 160G and the base 110 . The third filter element 162R is disposed between the third photo sensor 160R and the base 110 . The filter element can filter the excitation light before sensing. For example, the first filter element 162B can further filter out light other than the first excitation light EB to prevent the first photo sensor 160B from receiving the third light. The second excitation light EG and the third excitation light ER reduce the sensing accuracy. The second filter element 162G can further filter light other than the second excitation light EG to prevent the second photo sensor 160G from receiving the first excitation light EB and the third excitation light ER and thereby reducing the sensing accuracy. The third filter element 162R can further filter light other than the third excitation light ER to prevent the third photo sensor 160R from receiving the first excitation light EB and the second excitation light EG and thereby reducing the sensing accuracy.

The thermal convection polymerase chain reaction device 100 of this embodiment may further include a circuit board 170 . The base 110 and the heating element 130 can be disposed on the circuit board 170 , and the light combining element 140 can be disposed below the circuit board 170 . The first light-emitting element 150B, the second light-emitting element 150G and the third light-emitting element 150R can be electrically connected to the circuit board 170 .

In this embodiment, the base 110 has a plurality of elongated through holes 112 . The long axis A10 of the elongated through hole 112 is perpendicular to the long axis A20 of the tube to be tested 50 carried in the base 110 . The plurality of elongated through holes 112 can be connected to the first photo sensor 160B, the second photo sensor 160G and the third photo sensor 160R respectively to provide the first excitation light EB, The second excitation light EG and the third excitation light ER pass through. The elongated through hole 112 can produce a collimating effect on the first excitation light EB, the second excitation light EG and the third excitation light ER. The elongated through hole 112 is perpendicular to the tube 50 to be tested, which is equivalent to perpendicular to the incident light direction when the three colored lights illuminate the tube 50 to be tested. Therefore, the probability that the three colored lights enter the three sensors through the elongated through hole 112 can be greatly reduced, thereby reducing interference and improving sensing accuracy. In addition, in order to increase the amount of light input, a plurality of corresponding elongated through holes 112 can be provided for each sensor, so that the excitation light can be collimated through the elongated through holes 112 before sensing. As shown in FIG. 1 , in this embodiment, three elongated through holes 112 arranged along the long axis A20 of the tube to be tested 50 carried in the base 110 are correspondingly connected to the third sensor 160R to provide the first The excitation light EB, the second excitation light EG and the third excitation light ER pass through, and are filtered by the third filter element 162R before passing to the third sensor 160R.

In this embodiment, each light-emitting element may include a light-emitting diode and an optical filter. In detail, the first light-emitting element 150B includes a light-emitting diode 150B1 and an optical filter 150B2. The optical filter 150B2 is disposed between the light-emitting diode 150B1 and the light combining element 140 . The optical filter 150B2 can ensure that the first color light LB irradiated to the tube to be tested 50 has a preset wavelength to improve sensing accuracy. Similarly, the second light-emitting element 150G includes a light-emitting diode 150G1 and an optical filter 150G2. The optical filter 150G2 is disposed between the light-emitting diode 150G1 and the light combining element 140 . The third light-emitting element 150R includes a light-emitting diode 150R1 and an optical filter 150R2. The optical filter 150R2 is disposed between the light-emitting diode 150R1 and the light combining element 140 . The light emitting diode may be an organic or inorganic light emitting diode.

FIG. 3 is a schematic diagram illustrating the function of the light condensing unit of the thermal convection polymerase chain reaction device of FIG. 1 . Please refer to Figure 3. In this embodiment, the thermal convection polymerase chain reaction device 100 further includes a light condensing unit 180, which is disposed between the light emitting area R12 of the light combining element 140 and the base 110, and is suitable for converting the first color At least two of the light LB, the second color light LG and the third color light LR are focused on the tube to be tested 50, and are suitable for adjusting the focusing position. FIG. 4 is an adjusted schematic diagram of the light condensing unit of the thermal convection polymerase chain reaction device in FIG. 3 , and some components of the thermal convection polymerase chain reaction device are omitted in FIGS. 3 and 4 . In FIG. 3 , the light condensing unit 180 focuses the combined light of multiple colored lights (ie, at least two of the first colored light LB, the second colored light LG, and the third colored light LR) at a higher position on the tube to be tested 50 . On the other hand, in FIG. 4 , the light condensing unit 180 focuses the combined light of multiple colors of light at a lower position on the tube to be tested 50 . That is, adjusting the light condensing unit 180 can adjust the focused position. In the process of adjusting the focus position, you can find a good position for adjusting the focus, that is, a position where the three sensors have good sensing effects.

In addition, the focusing unit 180 can avoid the focusing position from the bottom of the tube 50 to be tested. Since impurities are easily deposited on the bottom of the tube to be tested 50 , avoiding the focus position from the bottom of the tube to be tested 50 can also reduce interference and improve sensing accuracy. The focusing position can be adjusted to be located at the upper half of the substance in the tube to be tested 50 and away from the bottom of the tube to be tested 50 . In this embodiment, the focusing unit 180 includes a lens barrel 182 and a focusing lens 184. The focusing lens 184 is movably disposed in the lens barrel 182, and the relative positions of the lens barrel 182 and the focusing lens 184 are moved. , you can adjust the focus position.

Figure 5 is a view of the light combining element of the thermoconvection polymerase chain reaction device of Figure 1 Exploded diagram. FIG. 6 is a partial top view of the light combining element of FIG. 5 . Please refer to FIGS. 5 and 6 . In this embodiment, the connected first plate-shaped part 140A, the second plate-shaped part 140B, the third plate-shaped part 140C and the fourth plate-shaped part 140D form an X-shaped structure. For example, the included angles between the first plate-like portion 140A, the second plate-like portion 140B, the third plate-like portion 140C and the fourth plate-like portion 140D are all 90 degrees, but the present disclosure is not limited thereto. In this embodiment, the light combining element 140 further includes two fixing seats 140E. The first plate-shaped part 140A, the second plate-shaped part 140B, the third plate-shaped part 140C and the fourth plate-shaped part 140D are fixed between the two fixing seats 140E.

For example, the first plate-shaped part 140A and the third plate-shaped part 140C may have the same optical properties, that is, the first plate-shaped part 140A and the third plate-shaped part 140C both reflect the first color light LB and allow the second color light LB to be reflected. The color light LG and the third color light LR pass through. The second plate-like part 140B and the fourth plate-like part 140D may have the same optical properties, that is, the second plate-like part 140B and the fourth plate-like part 140D both reflect the third color light LR and allow the second color light LG to interact with the first color light LR. The colored light LB passes through. Therefore, the first plate-shaped part 140A and the third plate-shaped part 140C of this embodiment may be the same optical element, while the second plate-shaped part 140B and the fourth plate-shaped part 140D are two separate optical elements. Such a design is only for convenience of assembly, but the present disclosure is not limited thereto.

In this embodiment, the wavelength of the first color light LB is, for example, between 451.5nm and 486.5nm, the wavelength of the second color light LG is, for example, between 507nm and 527nm, and the wavelength of the third color light LR is, for example, between 612nm and 644nm. . In another embodiment, the wavelength of the first color light LB is, for example, between 506 nm and 534 nm, the wavelength of the second color light LG is, for example, between 543 nm and 566 nm, and the wavelength of the third color light LR is, for example, If it is between 672nm and 696nm, the first excitation light EB generated by irradiating the material in the test tube 50 according to the color light range of the above interval is suitable for detecting influenza A virus, for example, and the second excitation light EG is suitable for detecting influenza type B virus, for example. , the third excitation light ER is suitable for detecting the Covid-19 virus, for example.

In summary, in the thermal convection polymerase chain reaction device and its optical detection method disclosed in the present disclosure, because a light combining element is used, multiple colors of light can be combined for detection, and multiple targets can be detected simultaneously, thereby reducing The amount of specimens used reduces the burden on the subjects.

50: Tube to be tested

60:Spectrometer

100: Thermal convection polymerase chain reaction device

110: base body

112: Slender through hole

130:Heating element

140: Light combining element

140A: First plate-shaped part

140B: Second plate-shaped part

140C: The third plate-shaped part

140D: The fourth plate-like part

150B: First light-emitting element

150G: Second light-emitting element

150R: The third light-emitting element

150B1,150G1,150R1: LED

150B2,150G2,150R2: Optical filter

160R: Third light sensor

162R: The third filter element

170:Circuit board

R12: light emission area

R14: first light entrance area

R16: Second light incident area

R18: The third light entrance area

LB: first color light

LG: Second color light

LR: third color light

ER: third excitation light

A10,A20: long axis

Claims (10)

A thermal convection polymerase chain reaction device, including: a base body suitable for carrying a tube to be tested; a heating element installed on the base body to heat the tube to be tested through the base body; a light combining element including a connected a first plate-like part, a second plate-like part, a third plate-like part and a fourth plate-like part, wherein the first plate-like part and the second plate-like part define a light emitting area, and the third plate-like part The two plate-shaped parts and the third plate-shaped part define a first light-incident area, the third plate-shaped part and the fourth plate-shaped part define a second light-incident area, and the first plate-shaped part and the fourth plate-shaped part The plate-shaped portion defines a third light-incident area; at least two light-emitting elements; at least two light sensors; and a light-condensing unit, which is disposed between the light-emitting area of the light-combining element and the base, and is suitable for integrating a The combined light of at least two of the first color light, a second color light and a third color light is focused on the tube to be tested, and is suitable for adjusting the height position of the focus on the tube to be tested, wherein the at least two lights The number of sensors corresponds to the number of the at least two light-emitting elements. The at least two light-emitting elements are at least two of a first light-emitting element, a second light-emitting element and a third light-emitting element. The first light-emitting element is suitable for When providing the first color light toward the first light incident area, the second light emitting element is adapted to provide the second color light toward the second light incident area, and the third light emitting element is adapted to provide the third light incident area toward the third light incident area. The third color light, the first plate-shaped portion is adapted to reflect the first color light and allow the second color light to interact with the third color light. Three-color light passes through, and the second plate-shaped part is adapted to reflect the third color light and allow the first color light and the second color light to pass, and the third plate-shaped part is adapted to reflect the first color light and allow the second color light to pass through. The colored light passes through, and the fourth plate-shaped portion is adapted to reflect the third colored light and allow the second colored light to pass. At least two of the first colored light, the second colored light and the third colored light enter the light emitting area and move towards The base emits light to illuminate the tube to be tested; and the at least two light sensors are at least two of a first light sensor, a second light sensor and a third light sensor, installed In the base, the first light sensor is adapted to receive a first excitation light generated by the substance in the tube to be tested being excited by the first color light, and the second light sensor is adapted to receive the The substance in the tube to be tested is excited by the second color light and generates a second excitation light. The third light sensor is adapted to receive the substance in the tube to be tested and is excited by the third color light and generates a third excitation light. excitation light. The thermal convection polymerase chain reaction device of claim 1, wherein the first plate-shaped part, the second plate-shaped part, the third plate-shaped part and the fourth plate-shaped part each have one surface connected to the other three are connected to form an X-shaped structure. The thermal convection polymerase chain reaction device as claimed in claim 1, wherein the focusing unit includes a lens barrel and a condenser lens, and the condenser lens is movably disposed in the lens barrel. The thermal convection polymerase chain reaction device according to claim 1, wherein the light synthesis element further includes two fixed seats, the first plate-shaped part, the second plate-shaped part, the third plate-shaped part and the third plate-shaped part. Four plate-shaped parts are fixed between the fixing seats. The thermal convection polymerase chain reaction device according to claim 1, further comprising at least two filter elements, the number of the at least two filter elements corresponding to the at least two light filters. The number of sensors, the at least two filter elements are at least two of a first filter element, a second filter element and a third filter element, wherein the first filter element is disposed on the third filter element between a light sensor and the base, the second filter element is disposed between the second light sensor and the base, and the third filter element is disposed between the third light sensor and the base between the seats. The thermal convection polymerase chain reaction device as claimed in claim 1, wherein the base body has a plurality of elongated through holes, and the long axes of the elongated through holes are perpendicular to the long axis of the tube to be tested carried in the base body, Correspondingly connected to the first light sensor, the second light sensor and the third light sensor. The thermal convection polymerase chain reaction device of claim 1, wherein each light-emitting element includes a light-emitting diode and a light filter, and the light filter is disposed between the light-emitting diode and the light combining element. between. An optical detection method is to use the thermal convection polymerase chain reaction device as described in claim 1 for detection, which includes: heating the substance in the tube to be tested; providing the first color light, the second color light and the At least two of the third color lights are combined, and the light combining element is used to combine the light. After the light is combined, the tube to be tested is irradiated; the light condensing unit is used to combine the first color light, the second color light and the third color light. The combined light of at least two of them is focused on the tube to be tested, and the focusing unit is adjusted to adjust the height position of the focus on the tube to be tested; and sensing that the substance in the tube to be tested is affected by the first color light The first excitation light generated by excitation, the second excitation light generated by excitation of the second color light and the third excitation light At least two of the third excitation lights are generated by exciting the colored light. The optical detection method according to claim 8, further comprising using at least two filter elements to filter at least two of the first excitation light, the second excitation light and the third excitation light before sensing. Test. The optical detection method of claim 8 further includes using a plurality of elongated through holes to collimate at least two of the first excitation light, the second excitation light and the third excitation light before sensing.

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