Multiple PCR (polymerase chain reaction) detection kit for anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris

A detection kit and technology for Anopheles, which is applied in the field of Anopheles beriorhynchus kits, multiplex PCR simultaneous detection of Anopheles spp. High and difficult to guarantee the accuracy of results, etc., to achieve the effect of cost and time saving, low requirements for personnel and equipment, and high safety in use

Active Publication Date: 2020-11-03
JIANGSU ACADEMY OF SCI & TECH FOR INSPECTION & QUARANTINE +2
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the identification of Anopheles mosquitoes mainly relies on morphological identification, and the accuracy of the results is difficult to guarantee, and for incomplete samples and samples in the non-adult stage, morphological identification cannot be performed; the method of molecular markers is time-consuming and costly, and has a great impact on personnel and equipment. The requirements are also higher

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  • Multiple PCR (polymerase chain reaction) detection kit for anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris
  • Multiple PCR (polymerase chain reaction) detection kit for anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris
  • Multiple PCR (polymerase chain reaction) detection kit for anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris

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Embodiment

[0032] 1. Screening test of primers for Anopheles mosquitoes. According to the conserved sequence of Anopheles mosquitoes gene (gene accession number MK685257.1), 6 sets of primers (groups 1-6) were designed:

[0033] Group 1:

[0034] F1: 5'-CAGCATGAGCAATTCCAGAT-3' (SEQ ID NO.1)

[0035] R1: 5'-GCAGGAACAGGATGAACTG-3' (SEQ ID NO.2)

[0036] Group 2:

[0037] F2: 5'-GCTAAATCTACTGATGCTCCA-3' (SEQ ID NO.3)

[0038] R2: 5'-TTTATCCTCCTCTTTCATCTGG-3' (SEQ ID NO.4)

[0039] Group 3:

[0040] F3: 5'-AGCATGAGCAATTCCAGAT-3' (SEQ ID NO.5)

[0041] R3: 5'-GGCAGGAACAGGATGAAC-3' (SEQ ID NO.6)

[0042] Group 4:

[0043] F4: 5'-GCATGTGCTGTTACAATAAC-3' (SEQ ID NO.7)

[0044] R4: 5'-CTGGTGCTTTCATTGGAGA-3' (SEQ ID NO.8)

[0045] Group 5

[0046] F5: 5'-GATGACCTAATTCAGCTCGA-3' (SEQ ID NO.9)

[0047] R5: 5'-GCCGGAATAGTAGGAACTTC-3' (SEQ ID NO.10)

[0048] Group 6:

[0049] F6: 5'-ATGACCTAATTCAGCTCGAA-3' (SEQ ID NO.11)

[0050] R6: 5'-AGCCGGAATAGTAGGAACT-3' (SEQ ID NO.12)

[0051] The P...

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Abstract

The invention relates to a multiple PCR (polymerase chain reaction) kit for simultaneously detecting anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris. The kit comprises specific primers for the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris, 10 * buffer, dNTPs, MgCl2, Taq enzymes, template DNA and sterile double distilled water, wherein the sequences of the specific primers for the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris are respectively shown as SEQ ID NO.5-6, SEQ ID NO.15-16 and SEQ ID NO.33-34. The multiple PCR detection kit disclosed by the invention can provide a qualitative detection result within 3 hours after a DNA sample is received, saves cost and time, has low requirements for personnel andequipment, and is an effective tool for detecting the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris.

Description

technical field [0001] The invention relates to a kit for simultaneous detection of Anopheles moltatus, Anopheles reckeii and Anopheles lanceolata by multiplex PCR. Background technique [0002] Anopheles belongs to the genus Anopheles of the insect class Diptera. There are about 450 known species and subspecies, which are divided into 6 subgenuses. They are distributed worldwide, mainly in tropical regions. Female Anopheles mosquitoes suck the blood of humans and animals, and can transmit diseases such as malaria parasites and filariasis. According to the report of the World Health Organization, there were 216 million cases of malaria in the world in 2016, and 445,000 people died of malaria and were affected by lymphatic filariasis. The number of people threatened was as high as 893 million, and about 40 million people were disfigured and lost their labor force by the disease. [0003] With the change of natural conditions and the development of international transportatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2537/143Y02A50/30
Inventor 杨庆贵陶莉何江周维田玲玲孙立新何德雨苏小建朱国强胡学锋贺骥谭伟龙
Owner JIANGSU ACADEMY OF SCI & TECH FOR INSPECTION & QUARANTINE
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