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A multiple PCR detection kit for Anopheles glabra, Anopheles multicolored and Anopheles aconitum

A detection kit, Anopheles aedae technology, applied in the Anopheles aconitum kit, multiplex PCR simultaneous detection of Anopheles aegypti, Anopheles multicolored, can solve the problem of high cost, difficult to guarantee the accuracy of results, personnel and High requirements for equipment and other issues, to achieve high specificity and sensitivity, cost and time savings, and low requirements for personnel and equipment

Active Publication Date: 2022-06-07
JIANGSU ACADEMY OF SCI & TECH FOR INSPECTION & QUARANTINE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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

Method used

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  • A multiple PCR detection kit for Anopheles glabra, Anopheles multicolored and Anopheles aconitum
  • A multiple PCR detection kit for Anopheles glabra, Anopheles multicolored and Anopheles aconitum
  • A multiple PCR detection kit for Anopheles glabra, Anopheles multicolored and Anopheles aconitum

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Embodiment

[0032] 1. Primer screening test of Anopheles meicolor, according to the conserved sequence of Anopheles meicolor gene (gene accession number MK685253.1), 6 groups of primers (numbered as 1-6 groups) were designed:

[0033] Group 1:

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

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

[0036] Group 2:

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

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

[0039] Group 3:

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

[0041] R3: 5'-CTTCCTCCCTCATTAACTTTAC-3' (SEQID NO.6)

[0042] Group 4:

[0043] F4: 5'-CTCCCGCATGAGCAATTC-3' (SEQID NO.7)

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

[0045] Group 5

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

[0047] R5: 5'-CTTGAGCTGGAATAGTAGGAA-3' (SEQID NO. 10)

[0048] Group 6:

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

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

[0051] The PCR rea...

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Abstract

The invention relates to a kit for simultaneously detecting Anopheles bechai, Anopheles multicolored and Anopheles aconitum by multiplex PCR. The kit includes specific primers for Anopheles glabra, Anopheles multicolored, and Anopheles aconitum, 10×buffer, dNTPs, MgCl 2 , Taq enzyme, template DNA, sterile double-distilled water, the sequences of the specific primers of Anopheles mechai, Anopheles multicolored, and Anopheles aconitum are respectively as SEQ ID NO.5-6, SEQ ID NO.13 -14, shown in SEQ ID NO.25-26. 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 Anopheles aconitum.

Description

technical field [0001] The invention relates to a kit for simultaneous detection of Anopheles mepicolor, Anopheles polyphemus and Anopheles aconitum by multiplex PCR. 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 changes in 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
CPCC12Q1/6888C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125Y02A50/30
Inventor 杨庆贵何江陶莉周有良谭伟龙苏小建邱文毅胡双双孙立新郭晔聂国嫒贺骥
Owner JIANGSU ACADEMY OF SCI & TECH FOR INSPECTION & QUARANTINE
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