One step method inverse transcription PCR kit for detecting and differentiating Zika viruses and detection method thereof

Abstract

The invention provides a primer, probe, and kit for detecting and differentiating Zika viruses through one step method inverse transcription PCR, a detection method and applications thereof. The PCR kit can be sensitively, specifically, efficiently, and rapidly detect and differentiate two genotypes of Zika viruses namely Asia type and Africa type at the same time. The principle of the kit is that on the basis of an inverse transcription PCR technology, a one step method FRET-PCR system, which can simultaneously detect and differentiate two Zika viruses, is established. The system is built on the basis of two genotypes of Zika viruses and whole gene sequences of other pathogens having a high homology with two genotypes of Zika viruses; a relatively conservative section is selected to design the primer and probe, through specific amplification, positive samples can be detected and screened from clinical samples sensitively and rapidly; then according to the difference of melting temperature (TM) values in a high resolution melting curve, the Asia type and Africa type of Zika viruses can be differentiated credibly; operation of the kit and the detection method is convenient and efficient, and the kit and method are suitable for detecting a large amount of samples.

Description

technical field [0001] The invention relates to the technical field of biological sciences, in particular to a kit for one-step reverse transcription PCR detection and typing of Zika virus and a detection method thereof. Background technique [0002] Zika virus disease, also known as Zika fever (Zika fever), is caused by Zika virus (Zika virus, ZIKAV) and passes through Aedes mosquitoes (mainly including Aedes aegypti, Aedes albopictus, Aedes africanus) Africana) and other acute viral vector-borne diseases transmitted by blood-sucking insects. The disease can also be transmitted from a pregnant woman to the fetus or infant during pregnancy or delivery, or through sexual transmission or blood transfusion. The incubation period of Zika virus infection is about 3-12 days. The main clinical symptoms are fever, rash, severe joint pain, muscle pain, and conjunctivitis. In cases of pediatric infection, changes in the nervous system, eyes, and hearing may also occur. Infection in p...

AI Technical Summary

Problems solved by technology

The technical solution of this patent application: 1) can only detect the Asian type of Zika virus; 2) it uses the fluorescent reverse transcription PCR of the hydrolysis probe (fluorescent oligonucleotide probe) for detection, so when performing the detection of related samples There are only amplification curves but no melting curves, so the negative and positive of the samples can only be judged by the presence or absence of the amplification curves, which cannot effectively reduce or avoid false positive results, and the feasibility of the detection results needs to be improved; 3) its kit The minimum detection limit is 1×10 3 copies / mL

Images