SFTSV detection kit using simplified nested fluorescent RT-PCR method

Abstract

The invention discloses a SFTSV detection kit using a simplified nested fluorescent RT-PCR method. The kit comprises two outer primersm, two internal primers and a probe. The DNA sequence of the first outer primer P1 is 5'-agt tca cag cag cat gga gag gat-3', the DNA sequence of the second outer primer P2 is 5'-agg ttg atg gca ctc cag gag aaa-3', the DNA sequence of the first internal primer P3 is 5'-agt tca cag cag cat gga gag gat-3', the DNA sequence of the second internal primer P4 is 5'-act ctc tgt ggc aag atg cct tca-3', and the DNA sequence of the probe is 5'-FAM-ttg ctg gct ccg cgc atc ttc aca tt-TAMRA-3'. Comparing with existing technologies, the kit possesses high specificity, sensitivity and repeatability in applications; the detection is rapid, simple and reliable; and the detecting time is shortened from 8 h to 3 h.

Description

technical field [0001] The invention relates to a test box for detecting SFTSV by a simple fluorescent RT-PCR method. Background technique [0002] In 2009, experts from the Chinese Center for Disease Control and Prevention isolated a new strain of bunyavirus from a patient's blood sample provided by Henan Province. In 2010, the Chinese Center for Disease Control and Prevention, through the monitoring of cases of unexplained fever with thrombocytopenia syndrome (SFTS) in many provinces, simultaneously discovered a new type of bunyavirus gene from the patient's serum by sequence-independent nucleic acid amplification technology According to the analysis of the whole genome information of the new bunyavirus and the analysis of electron microscope morphology, the new bunyavirus was determined to be a new virus belonging to the genus Phlebovirus in the family Bunyaviridae, named as fever with platelet Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV). Epidemiologi...

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Problems solved by technology

[0006] In view of the above-mentioned deficiencies, the technical problem to be solved in this invention is how to adapt to the requirements of the simple nested fluorescent RT-PCR proposed by integrating the advantages of the above-mentioned fluorescent RT-PCR and nested RT-PCR methods and removing its shortcomings, so that the The detection of SFTSV has the advantages of high sensitivity and specificity, non-pollution, simple operation, real-time rapidity, and good repeatability. A simple set of kits for the detection of SFTSV by fluorescent RT-PCR was designed.

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