HIV drug resistance detection carrier and construction method

Abstract

The invention relates to an HIV drug resistance detection carrier, and belongs to the technical field of genetic engineering. A construction method comprises the steps of performing first In-fusion connection on carriers pLWJ-SV40-Luc+ and LacZ genes so as to obtain a carrier pLWJ-SV40-Luc-LacZ, and performing second In-fusion connection on carriers pLWJ-SV40-Luc-LacZ and drug resistance detectiongenes so as to obtain an HIV drug resistance detection carrier, wherein the drug resistance detection genes are genes between 1678bp and 5042bp in an HIV virus Gag-Pol region in a sample to be detected. The HIV drug resistance detection carrier provided by the invention can detect the genes between the 1678bp and the 5042bp in the Gag-Pol region, including HIV-1 reverse transcriptase, protease and integrase inhibitors at the same time, and can cover main action target points of most majority of AIDS antivirus treatment medicines at present, and high-flux detection is realized.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an HIV drug resistance detection carrier and a construction method. Background technique [0002] With the widespread development of AIDS antiretroviral treatment, the emergence and importance of HIV drug resistance has become a focus of increasing attention, and it is urgent to prevent and control it. Fujian Province started free antiviral treatment in 2005. By the end of 2017, the province had provided free antiviral treatment to 10,299 patients. The results of drug resistance monitoring showed that patients who failed antiviral treatment in Fujian Province were resistant to NRTI, NNRTI and PI. Drug rates were 54.5% (72 / 132), 67.4% (89 / 132), 5.3% (7 / 132). HIV drug resistance has gradually become a major factor threatening the sustainable development of antiretroviral therapy. The International AIDS Association has recommended that viral drug resistance testing sho...

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Problems solved by technology

[0006] 2. The previously established phenotypic drug resistance detection method mainly uses enzyme-cut ligation cloning, which has low ligation efficiency, takes a long time, requires specific restriction enzyme sites, and is limited in application

However, this method only targets some protease and reverse transcriptase genes (1479bp), and cannot achieve high-throughput drug detection

Therefore, the existing phenotypic drug resistance detection kits have defects such as limited drug resistance detection range and low detection efficiency. High-throughput HIV-1 phenotypic detection reagents for all drug targets and their interactions

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