A multiplex PCR detection kit for An.

A detection kit and Anopheles technology, applied in the direction of recombinant DNA technology, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high cost, difficult to guarantee the accuracy of results, and high requirements for personnel and equipment. Achieve high specificity and sensitivity, save cost and time, and require less personnel and equipment

Active Publication Date: 2022-06-07
JIANGSU ACADEMY OF SCI & TECH FOR INSPECTION & QUARANTINE +2
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  • Abstract
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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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  • A multiplex PCR detection kit for An.
  • A multiplex PCR detection kit for An.
  • A multiplex PCR detection kit for An.

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Embodiment

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

[0033] Group 1:

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

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

[0036] Group 2:

[0037] F2: 5'-GCTAATCTACTGATGCTCCA-3' (SEQID NO.3)

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

[0039] Group 3:

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

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

[0042] Group 4:

[0043] F4: 5'-GCATGTGCTGTTACAATAAC-3' (SEQID 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' (SEQID NO.10)

[0048] Group 6:

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

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

[0051] The PCR...

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Abstract

The invention relates to a kit for simultaneous detection of Anopheles moltatus, Anopheles reckeii and Anopheles lanceolata by multiplex PCR. The kit includes specific primers for Anopheles papillonus, Anopheles reckeii, and Anopheles beriorhynchus, 10×buffer, dNTPs, MgCl 2 , Taq enzyme, template DNA, sterile double-distilled water, the sequences of the specific primers of Anopheles moltatus, Anopheles reckeii, and Anopheles lanceolata are respectively as SEQ ID NO.5-6, SEQ ID NO.15 -16, shown in SEQ ID NO.33-34. The multiplex PCR detection kit of the present invention can give qualitative detection results within 3 hours after receiving the DNA sample, saves cost and time, and has low requirements for personnel and equipment. and an effective tool for An.

Description

technical field [0001] The present invention relates to a multiplex PCR kit for simultaneously detecting Anopheles tadpoles, Anopheles reye and Anopheles barbatae. Background technique [0002] Anopheles belongs to the genus Anopheles, Diptera, Insecta. About 450 species and subspecies are known. They are divided into 6 subgenera and are distributed worldwide, mainly in tropical regions. The female Anopheles mosquitoes suck the blood of humans and animals, and can transmit diseases such as Plasmodium and filariasis. According to the World Health Organization report, in 2016, 216 million cases of malaria occurred worldwide, and 445,000 people died of malaria and were affected by lymphatic filariasis. The number of people threatened is as high as 893 million, and about 40 million people have been disfigured by disease and lost their workforce. [0003] With the change of natural conditions and the development of international transportation, trade and tourism, mosquitoes and ...

Claims

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Application Information

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Patent Type & Authority Patents(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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