CN110791570A - Primer group and kit for simultaneously detecting Wei-type, Si-type and heterodiscal paragonimiasis - Google Patents
Primer group and kit for simultaneously detecting Wei-type, Si-type and heterodiscal paragonimiasis Download PDFInfo
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Abstract
The invention discloses a primer group, a kit and a method for simultaneously detecting Wei-type, Si-type and heterodiscal paragonimiasis. Comprises a test strip, a DNA positive control solution of paragonimus and a detection solution, wherein the detection solution specifically comprises LAMP buffer solution, a primer group, Bst2.0DNA polymerase, dNTP and MgSO4And ddH2And O, the primer group comprises an outer primer F3, an outer primer B3, an inner primer FIP, an inner primer BIP and a Probe. The invention can quickly detect the paragonimus genomic DNA in a sample, provides a method for detecting the paragonimus genomic DNA, and can be used for detecting the genomic DNA of paragonimus adults, cysticercus and worm eggs. The invention has high sensitivity, strong specificity, better stability and simple operation, can better meet the requirements of personnel at different levels and is easy to popularize to the basic level.
Description
Technical Field
The invention belongs to the diagnostic technology in the field of biotechnology, and particularly relates to a primer for detecting Wei-type, Sishi-type and isoperimular paragonimus and application thereof.
Background
Adults of the paragonimus species colonize the lungs of the host, commonly known as the lung fluke. To date, nearly 50 or more paragonimus have been reported all over the world, 32 of which are reported in China, and paragonimus westermani, paragonimus minor and isoperipherus paragonimus are most prevalent and all can parasitize human bodies. The paragonimus latifolius adults often parasitize the lung, but the juvenile worms sometimes parasitize tissues and organs such as the liver, the brain, the eye socket, the subcutaneous tissue and the like, and various tissues and organs are damaged due to migration of the worms in the body. The paragonimiasis is complicated in symptoms, often involves multiple organs, brings a difficult problem to diagnosis of the paragonimiasis, and is often misdiagnosed as tuberculosis, pneumonia and the like.
The traditional paragonimiasis diagnostic methods include etiological diagnosis and immunological diagnosis. The method is characterized in that the method comprises the steps of (1) checking worm eggs with phlegm or excrement and biopsy of subcutaneous tissues in etiological diagnosis, and then the worm eggs are difficult to observe and easy to miss detection in a latent period or chronic infection; an intradermal test or an enzyme-linked immunosorbent assay, can be used for diagnostic tests, but cannot distinguish between a previous infection and a present infection. The invention designs a group of primers capable of simultaneously detecting paragonimus westermani, paragonimus schlegeli and isoperiphera paragonimus, combines a nucleic acid detection test strip, can be used for carrying out on-site rapid detection, does not need special instruments and equipment, is simple to operate, and has important significance for preventing and controlling diseases.
Disclosure of Invention
In order to overcome the defects in the prior detection technology and meet the requirements on disease prevention and control, the invention provides a primer group, a kit and a method for simultaneously detecting paragonimus westermani, paragonimus westermani and isoperimulus paragonimus, wherein the primer group, the kit and the method are used for performing loop-mediated isothermal amplification and have strong specificity and high sensitivity.
The invention provides a body primer group, a nucleic acid detection test strip and a detection solution, which specifically comprise LAMP buffer solution, 8U Bst2.0DNA polymerase, 1.5mM dNTP and 7.5mM MgSO 24And ddH2And O. The invention also provides a method for detecting the genomic DNA of the paragonimus, which can be used for detecting the genomic DNA of the paragonimus adults, cysticercus and ova, can accurately and quickly detect samples on site and is convenient to operate.
The technical scheme adopted by the invention is as follows:
a primer group capable of simultaneously detecting Wei-type, Siqi-type and heterodiscal paragonimus:
the specific sequences of the primer group are as follows:
outer primer F3: GCTTCGAATAGTCAGGGCG, as shown in SEQ ID No. 1;
outer primer B3: GCAGTGATACCAGCAGTGA, as shown in SEQ ID No. 2;
inner primer FIP: CTCGAACGCTGCGTCGTTGAGTAAGTGCTCAAGTTGCCA CC, as shown in SEQ ID No. 3;
the inner primer BIP: ACGTTCGACTACCCCCATTCGACGGATTAGCGTTCGATG GC, as shown in SEQ ID No. 4;
probe Probe: CAGGGTCACGTGATTCTCATCAGC, as shown in SEQ ID No. 5.
The concentrations of the outer primers F3 and B3 are both 0.2 mu mol/mu l, the concentrations of the inner primers FIP and BIP are both 1.6 mu mol/mu l, and the concentration of the Probe is 0.8 mu mol/mu l.
The 5 'end of the inner primer FIP in the primer group is modified by the biotin, and the 5' end of the Probe is modified by the FITC.
Secondly, a kit capable of simultaneously detecting Wei-type, Si-type and heterodiscal paragonimus:
comprises a nucleic acid detection test strip, a paragonimus DNA positive control solution and a detection solution, wherein the detection solution specifically comprises LAMP buffer solution, a primer group, Bst2.0DNA polymerase, dNTP and MgSO4And ddH2O, wherein the primer set is the primer set according to any one of claims 1 to 3.
The Bst2.0DNA polymerase, dNTP and MgSO4The concentrations in the detection solution were 8U, 1.5mM, and 7.5mM, respectively.
The LAMP buffer solution comprises Tris-HCl with the final concentration of 20mM, KCl with the final concentration of 10mM and (NH) with the final concentration of 10mM4)2SO4Tween-20 with the mass fraction of 0.1 percent and 1M betaine.
The concentration of the DNA of the paragonimus aquaticus in the DNA positive control solution is 50 ng/. mu.L.
The nucleic acid detection test strip comprises a sample pad, a nitrocellulose membrane and a water absorption pad which are sequentially arranged, wherein the water absorption pad is used as a handheld part; the nitrocellulose membrane is used as a reaction area, a gold label pad, a line C and a line T are arranged on the nitrocellulose membrane, the line C is a quality control line, the line T is a detection line, and the gold label pad, the line T and the line C are sequentially arranged on the nitrocellulose membrane from the sample pad to the water absorption pad; and (3) spraying anti-FITC (fluorescein isothiocyanate) labeled latex particles (the concentration is 1mg/mL) on the gold-labeled pad, wherein the T line is avidin (the concentration is 1mg/mL), and the C line is goat anti-mouse monoclonal antibody (the concentration is 1 mg/mL).
And thirdly, the application of the primer group or the kit for simultaneously detecting the Wei-type, the Si-type and the heterodiscal paragonimus:
the kit is used for detecting the genome DNA of adults, cysticercus and ova of Wei-, Si-and Heteropano-trematodes.
The use method of the kit for simultaneously detecting the Wei-type, the Sichuan-type and the heterodiscal paragonimus comprises the following steps:
the invention also provides a using method of the detection kit, which comprises the following steps:
1) and (3) detection procedures: extracting genome DNA from a sample to be detected, mixing the genome DNA with a detection solution, reacting in a water bath kettle at 63 ℃ for 45 minutes, adding a reaction product onto a test strip, observing the result after 5-10 minutes, and judging on the test strip for sample detection in the following way:
when the T line and the C line can be clearly seen, the T line and the C line are red, the lines are single, clear and light in background color, the detection result of the serum sample to be detected is judged to be positive, and the positive represents that the genomic DNA of the Wei-type, the Si-type or the heterodiscal paragonimus is detected;
when the T line cannot be clearly seen but the C line can be clearly seen and the C line is red, the detection result of the serum sample to be detected is judged to be negative, and the negative represents that the Wei-type, the Si-type or the heterodiscal paragonimus are not detected;
and when the T line and the C line cannot be clearly seen, namely the T line and the C line are not red, judging that the detection result of the serum sample to be detected is invalid, wherein the invalidity represents that the test strip is invalid, and replacing the test strip for redetection.
2) Mixing the paragonimus DNA positive control solution with the detection solution, reacting in a water bath kettle at 63 ℃ for 45 minutes, adding a reaction product onto a test strip, observing the result after 5-10 minutes, and judging on the test strip for sample detection in the following way:
when the T line and the C line can be clearly seen, the T line and the C line are red, the lines are single, clear and light in background color, and the positive control solution, the kit detection solution and the test strip can be used for normal detection and have accurate results;
when the T line cannot be clearly seen but the C line can be clearly seen, and the C line is red, the positive control solution is invalid;
and if the T line and the C line cannot be clearly seen, namely the T line and the C line are not red, the positive control solution, the detection solution of the kit or the test strip is invalid, and the detection cannot be accurately carried out.
The kit can be used for quickly detecting the paragonimus genomic DNA in the sample, has the advantages of high sensitivity, strong specificity, better stability and simple operation, can better meet the requirements of personnel at different levels, and is easy to popularize to the basic level.
The invention also provides a test strip based on nucleic acid detection, which can accurately and rapidly detect a sample on site and is convenient to operate.
Compared with the prior art, the paragonimus detection method has the following beneficial effects:
firstly, the primer group designed in the invention can simultaneously detect Weishi, Sishi and heterodiscal concurrent trematodes, has obvious advantages and innovation compared with the prior art, and is an important supplement for the detection of the disease;
thirdly, the LAMP reaction components in the invention are optimized, so that the specificity and the sensitivity are good;
thirdly, the probe and the modified primer are added, and the nucleic acid detection test strip is combined, so that the visualization, rapidness and simplification of the detection result are realized, and the kit has important practical significance for diagnosis and prevention of paragonimiasis.
Drawings
FIG. 1 shows the real-time fluorescence LAMP detection results, 1-8 are paragonimus westermani, paragonimus schlegelii, paragonimus dissimi, trichinella, hemiptera, schistosome, angiostrongylus cantonensis genome DNA and distilled water, respectively.
FIG. 2 is a schematic diagram of test strip result determination.
FIG. 3 shows the result of LAMP reaction product detection by test paper, 1-8 are LAMP reaction products using paragonimus westermani, paragonimus dorsalis, paragonimus isoperiformis, trichina, anisakis, schistosome, angiostrongylus cantonensis genome DNA and distilled water as templates, respectively.
Detailed Description
The present technology is further described in conjunction with the appended drawings and the specific embodiments, it is to be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
The examples of the invention are as follows:
example 1 real-time fluorescent LAMP validation of primer effectiveness
Primers F3, B3, FIP and BIP, 8U Bst2.0DNA polymerase, 1.5mM dNTP, 7.5mM MgSO 2 were added to the reaction tube4And ddH2O, wherein the concentrations of the outer primers F3 and B3 are both 0.2. mu. mol/. mu.l, the concentrations of the inner primers FIP and BIP are both 1.6. mu. mol/. mu.l, and 1. mu.l of SYTO13 fluorescent dye with a concentration of 25. mu.M is further added, and 2. mu.l of Wei's, Sichuan and Heteropanus paragonimus adult genomic DNAs obtained by extraction with a Tiangen blood/cell/tissue genomic DNA extraction kit (cat No. DP304) are added as templates, respectively. The reaction is carried out on a fluorescence quantitative PCR instrument of BerleThe specific reaction procedure is as follows: react at 65 ℃ for 1.5h and collect fluorescence data, and react at 95 ℃ for 5 min. The detection results are shown in figure 1, the primer can simultaneously detect the genomic DNA of Wei's, Sis and Heteropanus paragonimus adults, and can not detect the genomic DNA of trichina, anisakis, schistosome and angiostrongylus cantonensis, which indicates that the primer group has good specificity.
Example 2 detection of LAMP reaction product by nucleic acid detection test strip
Adding the LAMP product after reaction into a test strip, waiting for 5-10 minutes, clearly seeing a T line and a C line, judging that the detection result of the sample is positive if the lines are single, clear and light in background color; in the test paper strip for sample detection, the T line cannot be clearly seen, but the C line is clearly red, the detection result of the sample is judged to be negative, and the T line and the C line are not red, so the detection result is invalid. The results are illustrated in FIG. 2. The test results are shown in fig. 3, in which the test strips of LAMP reaction products with templates of genomic DNA of Euonymus, Strongyloides and Heteropana were positive, while the test strips with templates of genomic DNA of Trichinella spiralis, Heterodera anisum, Schistosoma japonicum, angiostrongylus cantonensis or distilled water were negative.
Example 3 test results of clinical samples with the kit
The kit is used for detecting 124 canine blood samples, genome DNA is extracted from the blood samples to be detected, 5 mu l of the extracted genome DNA is added into LAMP reaction liquid to serve as a template, the LAMP reaction liquid reacts for 45 minutes in a water bath at 63 ℃, reaction products are added onto test paper strips, and the results are observed after 5-10 minutes. The results showed that 9 positive samples were detected with a positive rate of 7.25%, 115 negative samples with a negative rate of 92.75%, and the results are shown in Table 1.
Table 1 test strip clinical test results
| Positive (%) | Negative (%) | |
| Percentage of detection (%) | 9(7.25) | 115(92.75) |
And (4) judging a result: in the test strip for sample detection, a T line and a C line can be clearly seen, the lines are single and clear, and the background color is light, so that the detection result of the sample is judged to be positive paragonimiasis infection; in the test paper strip for sample detection, the T line cannot be clearly seen, but the C line is clearly red, the detection result of the sample is judged to be negative to paragonimus infection, and the T line and the C line are not red, so the detection result is invalid.
On the premise that all the test strips can clearly see the C line, clear red T lines and red C lines can be seen on the test strips of the positive standard, clear C lines can be seen but the T lines cannot be seen on the test strips of the negative standard, the lines are single, clear and light in background color, the result of sample detection can be judged, and otherwise, the result is redone.
While the invention has been described in connection with a preferred embodiment, it will be understood that various changes and modifications may be effected therein by one skilled in the art after reading the foregoing description, and equivalents may be resorted to, falling within the scope of the invention as defined by the appended claims.
The gene sequence involved in the invention is as follows:
SEQ ID No.1；
name: outer primer F3 gene sequence
The source is as follows: artificially synthesized
GCTTCGAATAGTCAGGGCG
SEQ ID No.2；
Name: outer primer B3 gene sequence
The source is as follows: artificially synthesized
GCAGTGATACCAGCAGTGA
SEQ ID No.3；
Name: inner primer FIP gene sequence
The source is as follows: artificially synthesized
CTCGAACGCTGCGTCGTTGAGTAAGTGCTCAAGTTGCCACC
SEQ ID No.4；
Name: BIP gene sequence of inner primer
The source is as follows: artificially synthesized
ACGTTCGACTACCCCCATTCGACGGATTAGCGTTCGATGGC
SEQ ID No.5；
Name: probe gene sequence
The source is as follows: artificially synthesized
CAGGGTCACGTGATTCTCATCAGC。
Sequence listing
<110> Zhejiang province academy of medical science
<120> primer group and kit for simultaneously detecting Wei-type, Sishi and heterodiscal paragonimiasis
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<170>SIPOSequenceListing 1.0
<210>1
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>1
gcttcgaata gtcagggcg 19
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<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>2
gcagtgatac cagcagtga 19
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<212>DNA
<213> Artificial Sequence (Artificial Sequence)
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ctcgaacgct gcgtcgttga gtaagtgctc aagttgccac c 41
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<213> Artificial Sequence (Artificial Sequence)
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acgttcgact acccccattc gacggattag cgttcgatgg c 41
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<212>DNA
<213> Artificial Sequence (Artificial Sequence)
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cagggtcacg tgattctcat cagc 24
Claims (9)
1. A primer group capable of simultaneously detecting Wei-type, Si-type and Bidiscal paragonimus is characterized in that:
the specific sequences of the primer group are as follows:
outer primer F3: GCTTCGAATAGTCAGGGCG, as shown in SEQ ID No. 1;
outer primer B3: GCAGTGATACCAGCAGTGA, as shown in SEQ ID No. 2;
inner primer FIP: CTCGAACGCTGCGTCGTTGAGTAAGTGCTCAAGTTGCCA CC, as shown in SEQ ID No. 3;
the inner primer BIP: ACGTTCGACTACCCCCATTCGACGGATTAGCGTTCGATG GC, as shown in SEQ ID No. 4;
probe Probe: CAGGGTCACGTGATTCTCATCAGC, as shown in SEQ ID No. 5.
2. The primer set for simultaneous detection of Fangwei, Siwei and Mesordidae according to claim 1, wherein: the concentrations of the outer primers F3 and B3 are both 0.2 mu mol/mu l, the concentrations of the inner primers FI P and BIP are both 1.6 mu mol/mu l, and the concentration of the Probe is 0.8 mu mol/mu l.
3. The primer set for simultaneous detection of Fangwei, Siwei and Mesordidae according to claim 1, wherein: the 5 'end of the inner primer FIP in the primer group is modified by the biotin, and the 5' end of the Probe is modified by the FITC.
4. The utility model provides a can detect wei formula, si formula and heterodiscal coculture worm's kit which characterized in that: comprises a test strip, a DNA positive control solution of paragonimus and a detection solution, wherein the detection solution specifically comprises LAMP buffer solution, a primer group, Bst2.0DNA polymerase, dNTP and MgSO4And ddH2O, wherein the primer set is the primer set according to any one of claims 1 to 3.
5. The primer set and the kit for simultaneously detecting Wei-, Si-and Bisbon-worms according to claim 4, wherein: the LAMP buffer solution comprises Tris-HCl with the final concentration of 20mM, KCl with the final concentration of 10mM and (NH) with the final concentration of 10mM4)2SO4Tween-20 with the mass fraction of 0.1 percent and 1M betaine.
6. The primer set and the kit for simultaneously detecting Wei-, Si-and Bisbon-worms according to claim 4, wherein: the concentration of the DNA of the paragonimus aquaticus in the DNA positive control solution is 50 ng/. mu.L.
7. The primer set and the kit for simultaneously detecting Wei-, Si-and Bisbon-worms according to claim 4, wherein: the test strip comprises a sample pad, a nitrocellulose membrane and a water absorption pad which are sequentially arranged, wherein the water absorption pad is used as a handheld part; the nitrocellulose membrane is used as a reaction area, a gold label pad, a line C and a line T are arranged on the nitrocellulose membrane, the line C is a quality control line, the line T is a detection line, and the gold label pad, the line T and the line C are sequentially arranged on the nitrocellulose membrane from the sample pad to the water absorption pad; and (3) spraying anti-FITC (fluorescein isothiocyanate) labeled latex particles on the gold label pad, wherein the T line is avidin, and the C line is goat anti-mouse monoclonal antibody.
8. Use of the primer set for simultaneous detection of a Euonymus, Sjogren and paragonimus according to any one of claims 1 to 3 or the kit for simultaneous detection of a Euonymus, Sjogren and paragonimus according to any one of claims 4 to 7, wherein: the kit is used for detecting the genome DNA of adults, cysticercus and ova of Wei-, Si-and Heteropano-trematodes.
9. Use of a kit for the simultaneous detection of a Euonymus, Strongyloides and Parazoea according to any one of claims 4 to 7, characterized in that it comprises:
1) and (3) detection procedures: extracting genome DNA from a sample to be detected, mixing the genome DNA with a detection solution, reacting in a water bath kettle at 63 ℃ for 45 minutes, adding a reaction product onto a test strip, observing the result after 5-10 minutes, and judging on the test strip for sample detection in the following way:
when the T line and the C line can be clearly seen, the T line and the C line are red, the lines are single, clear and light in background color, the detection result of the serum sample to be detected is judged to be positive, and the positive represents that the genomic DNA of the Wei-type, the Si-type or the heterodiscal paragonimus is detected;
when the T line cannot be clearly seen but the C line can be clearly seen and the C line is red, the detection result of the serum sample to be detected is judged to be negative, and the negative represents that the Wei-type, the Si-type or the heterodiscal paragonimus are not detected;
and when the T line and the C line cannot be clearly seen, namely the T line and the C line are not red, judging that the detection result of the serum sample to be detected is invalid, wherein the invalidity represents that the test strip is invalid, and replacing the test strip for redetection.
2) Mixing the paragonimus DNA positive control solution with the detection solution, reacting in a water bath kettle at 63 ℃ for 45 minutes, adding a reaction product onto a test strip, observing the result after 5-10 minutes, and judging on the test strip for sample detection in the following way:
when the T line and the C line can be clearly seen, the T line and the C line are red, the lines are single, clear and light in background color, and the positive control solution, the kit detection solution and the test strip can be used for normal detection and have accurate results;
when the T line cannot be clearly seen but the C line can be clearly seen, and the C line is red, the positive control solution is invalid;
and if the T line and the C line cannot be clearly seen, namely the T line and the C line are not red, the positive control solution, the detection solution of the kit or the test strip is invalid, and the detection cannot be accurately carried out.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111719004A (en) * | 2020-08-06 | 2020-09-29 | 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心) | Kit for distinguishing and identifying paragonimus westermani cysticercus and paragonimus cysticercus and using method |
| CN111808973A (en) * | 2020-08-06 | 2020-10-23 | 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心) | Kit for identifying tripartite paragonium metacercaria and using method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980028711A (en) * | 1996-10-24 | 1998-07-15 | 박원훈 | Gene Sequences and Recombinant Proteins Encoding Cysteine Proteases in Pulmonary Insects |
| CN101266253A (en) * | 2008-04-25 | 2008-09-17 | 中国人民解放军第三军医大学 | Paragonimiasis detection reagent kit and method for making same |
| CN101266252A (en) * | 2008-04-25 | 2008-09-17 | 中国人民解放军第三军医大学 | Paragonimiasis detection device and method for making same |
| CN101339190A (en) * | 2008-05-19 | 2009-01-07 | 浙江省医学科学院 | Human body important parasite antigen chip and method for making same |
| CN101956013A (en) * | 2010-09-29 | 2011-01-26 | 中国疾病预防控制中心寄生虫病预防控制所 | Kit and method for detecting paragonismus westermani quickly |
| CN102279259A (en) * | 2010-06-10 | 2011-12-14 | 中国疾病预防控制中心寄生虫病预防控制所 | Diagnostic reagent kit for paragonimiasis detection and preparation method thereof |
-
2019
- 2019-10-30 CN CN201911045773.0A patent/CN110791570B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980028711A (en) * | 1996-10-24 | 1998-07-15 | 박원훈 | Gene Sequences and Recombinant Proteins Encoding Cysteine Proteases in Pulmonary Insects |
| CN101266253A (en) * | 2008-04-25 | 2008-09-17 | 中国人民解放军第三军医大学 | Paragonimiasis detection reagent kit and method for making same |
| CN101266252A (en) * | 2008-04-25 | 2008-09-17 | 中国人民解放军第三军医大学 | Paragonimiasis detection device and method for making same |
| CN101339190A (en) * | 2008-05-19 | 2009-01-07 | 浙江省医学科学院 | Human body important parasite antigen chip and method for making same |
| CN102279259A (en) * | 2010-06-10 | 2011-12-14 | 中国疾病预防控制中心寄生虫病预防控制所 | Diagnostic reagent kit for paragonimiasis detection and preparation method thereof |
| CN101956013A (en) * | 2010-09-29 | 2011-01-26 | 中国疾病预防控制中心寄生虫病预防控制所 | Kit and method for detecting paragonismus westermani quickly |
Non-Patent Citations (3)
| Title |
|---|
| YOUNG-AN BAE ET AL: "PwRn1, a novel Ty3/gypsy-like retrotransposon of Paragonimus westermani: molecular characters and its differentially preserved mobile potential according to host chromosomal polyploidy", 《BMC GENOMICS》 * |
| ZHUO XUNHUI ET AL: "DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick", 《ACTA TROPICA》 * |
| 赵昕等: "卫氏并殖吸虫P C R 和实时荧光PPCR快速检测方法的建立", 《生物技术通报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111719004A (en) * | 2020-08-06 | 2020-09-29 | 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心) | Kit for distinguishing and identifying paragonimus westermani cysticercus and paragonimus cysticercus and using method |
| CN111808973A (en) * | 2020-08-06 | 2020-10-23 | 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心) | Kit for identifying tripartite paragonium metacercaria and using method |
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