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Multiple PCR detection kit for Anopheles splendidus, Anopheles maculatus and Anopheles aconitus

A detection kit, Anopheles mosquito technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of molecular labeling methods that take a long time, high requirements for personnel and equipment, and accurate results. It can save costs and time, reduce the requirements for personnel and equipment, and achieve high safety in use.

Active Publication Date: 2020-09-22
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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  • Multiple PCR detection kit for Anopheles splendidus, Anopheles maculatus and Anopheles aconitus
  • Multiple PCR detection kit for Anopheles splendidus, Anopheles maculatus and Anopheles aconitus
  • Multiple PCR detection kit for Anopheles splendidus, Anopheles maculatus and Anopheles aconitus

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Experimental program
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Embodiment

[0032] 1. Primer screening test for Anopheles acanthus, based on the conserved sequence of the Anopheles acanthus gene (gene accession number MK685253.1), designed 6 sets of primers (groups 1-6):

[0033] Group 1:

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

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

[0036] Group 2:

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

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

[0039] Group 3:

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

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

[0042] Group 4:

[0043] F4: 5'-CTCCCGCATGAGCAATTC-3' (SEQ ID 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' (SEQ ID NO.10)

[0048] Group 6:

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

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

[0051] The PCR reaction soluti...

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Abstract

The invention relates to a kit for simultaneous detection of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus by multiple PCR. The kit includes specific primers of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus, 10xbuffer, dNTPs, MgCl2, Taq enzyme, template DNA and sterile double distilled water, wherein sequences of the specific primers of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus are shown as SEQ ID NO.5-6, SEQ ID NO. 13-14, and SEQ ID NO 25-26. The multiple PCR detection kit can provide qualitative detection results within 3 hours after receiving DNA samples, the cost and time are saved, the requirements on personnel and equipment are low, and the multiple PCR detection kit is an effective tool for detecting Anopheles splendidus,Anopheles maculatus and Anopheles aconitus.

Description

technical field [0001] The invention relates to a kit for simultaneously detecting Anopheles bechai, Anopheles multicolored and Anopheles aconitum 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
CPCC12Q1/6888C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125Y02A50/30
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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