Enterovirus triple real-time fluorescent quantitative RT-PCR detection kit

Abstract

The invention provides an enterovirus triple real-time fluorescent quantitative RT-PCR detection kit, which comprises a quantitative RT-PCR reaction solution, a standard substance and a reference substance, wherein, the quantitative RT-PCR reaction solution comprises a RT-PCR buffer solution, MgCl2, dNTPs, an EV universal primer, an EV universal fluorescent probe, an EV71 type fluorescent probe, a CA16 type fluorescent probe, heat resistant DNA polymerase and reverse transcriptase. The kit of the invention enables detection and genotyping to enterovirus universal type, enterovirus 71 type andcoxsackie virus group A 16 type by a one-step method. Compared with the current single detection kit, the kit of the invention is capable of saving the production cost and detection cost, raising theefficiency and time of the detection. Under the condition of ensuring no competition of multiple primers, the purpose of multi-channel detection can be achieved, so that the unification of the efficiency and technology is achieved.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a fluorescent quantitative RT-PCR detection kit, in particular to a three-probe real-time fluorescent quantitative RT-PCR for detection of enteroviruses in children's feces, pharynx, blood, cerebrospinal fluid and other samples. (EV), enterovirus type 71 (EV71), and coxsackievirus A16 (CA16) typing and quantification. This product takes the highly conserved 5' non-coding region (UTR) of enteroviruses as the target sequence, and designs enterovirus general primers and three-color fluorescent markers for enterovirus general type, enterovirus 71 and coxsackie virus A16 The probe uses one-step three-color real-time fluorescent quantitative RT-PCR technology to achieve the application purpose of early, rapid and accurate detection and typing of all enteroviruses. Background technique [0002] Hand-foot-mouth disease (HFMD) is a common infectious disease caused by a variety of enter...

AI Technical Summary

Problems solved by technology

Hand, foot and mouth disease caused by the two is difficult to distinguish clinically

Traditional enterovirus detection includes virus isolation and culture and serological diagnosis. The former was once regarded as the "gold standard" for enterovirus diagnosis, but the operation is cumbersome and time-consuming, generally takes 4-15 days, and there are cumbersome and sensitive operations. It is far from meeting the "four early" requirements of disease prevention; and the most commonly used serological diagnosis is the neutralization test, that is, the microplate method is used to determine the antibody titer, and the titer of the serum in the acute phase and the serum in the convalescent phase are usually used for testing. In comparison, a 4-fold or more increase in antibody titers proves viral infection

However, it can only be used as a basis for retrospective diagnosis, and it is easy to cross-react with other viruses, so the antibody method is not helpful for early diagnosis

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