B19 (Human parvovirus B19), HTLV (Human T-cell lymphotropic virus) and HEV (Hepatitis E virus) quadruple fluorescent PCR (polymerase chain reaction) quick hypersensitivity detection kit and application thereof

Abstract

The invention provides a B19 (Human parvovirus B19), HTLV (Human T-cell lymphotropic virus) and HEV (Hepatitis E virus) quadruple fluorescent PCR (polymerase chain reaction) quick hypersensitivity detection kit and an application thereof, and relates to a real-time PCR method for quadruple detection of target nucleic acid in a nucleic acid extraction liquid in a single PCR reaction container. The method is used for detecting B19, HTLV-I, HTLV-2 and HEV.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection. Specifically, the invention relates to the ability to simultaneously detect human parvovirus B19 (Human parvovirus B19, B19), human T-cell lymphotropic virus type I (Human T-cell lymphotropic virus 1, HTLV-I), A multiplex real-time PCR method for detecting four pathogenic pathogens of human T-cell lymphotropic virus 2 (HTLV-II) and hepatitis E virus (HEV), which is fast and ultrasensitive And complete in one step. In addition, the present invention also relates to the reagents involved in the above PCR method, such as primers, probes, etc. that are irrelevant to each other, as well as the detection kit used in the above method and the preparation and application of the corresponding detection kit. Background technique [0002] Human parvovirus B19 (Human parvovirus B19) is a linear single-stranded DNA virus with a genome size of 5.6 kb. In 1975, when Cossart et al. detected the h...

AI Technical Summary

Problems solved by technology

[0011] Although human parvovirus B19 (Human parvovirus B19), human T-cell lymphotropic virus type I (Human T-cell lymphotropic virus 1, HTLV-I), human T-cell lymphotropic virus type II (Human T-cell lymphotropic virus 2, HTLV- II), hepatitis E virus (Hepatitis E virus, HEV) these four kinds of viruses, these viruses themselves all can use PCR method to carry out independent detection respectively, but their joint detection has not reported yet

At present, conventional real-time PCR detection, such as Chinese patents CN1768151A, CN101189505A, CN101273262A, CN101302473A, etc., are mostly only used to detect a single target nucleic acid

The Chinese patent CN101624629A of the inventor team discloses (in a single PCR reaction container) a real-time PCR method for multiple detection of target nucleic acids in samples, which is faster and more integrated, but it is only used to detect specific multiple target nucleic acids detection, the platform technology cannot be extended to other virus combinations without creative labor

[0012] Especially at present, the nucleic acid multiplex detection technology industry mostly keeps its own platform technology secret, and there is no hint on how to design primer pairs and probes that do not interfere with each other, let alone adjust the concentration of the probe

The inventors have found that the quadruple primer pairs and probes that do not interfere with each other cannot be designed directly through the currently known primer pair and probe sequence design methods (including existing bioinformatics software), and the designed will Cause mutual interference and distort the fluorescence detection curve

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