Human enterovirus four-color fluorescence RT-PCR detection kit and detection method

Abstract

The invention discloses a human enterovirus four-color fluorescence RT-PCR detection kit and a detection method; the detection kit comprises multiple fluorescence RT-PCR reaction mother liquor, AMV reverse transcriptase liquid, Taq DNA polymerase liquid, a positive control, and a negative control; the detection kit is characterized by further comprising an internal control, a multiple 10*PCR reaction buffer, bovine serum albumin with a concentration of 20 mg / ml, various forward primers and reverse primers with a concentration of 100 muM, and various specific probes with a concentration of 50 muM, wherein sequences of the forward primers, reverse primers, specific probes and the internal control are as shown in SEQIDNO. 1, NO. 2, NO.3, NO.4, NO.5, NO.6, NO.7. NO.8, NO.9, NO.10., NO.11., NO.12., and NO.13. The detection kit of the invention has the advantages of high detection sensitivity, good specificity, accuracy, high speed, and no false positivity.

Description

technical field [0001] The invention relates to a fluorescent PCR kit in the field of biotechnology, in particular to a four-color fluorescent RT-PCR detection kit containing an internal control for general detection of human enteroviruses and type detection of enteroviruses EV71 and CoxA16 . Background technique [0002] Enteroviruses include 71 serotypes including poliovirus, Coxsackievirus, enterocytopathic human orphan virus (ECHO) and new enteroviruses. The hand, foot and mouth disease caused by enterovirus is a kind of acute infectious disease. The main clinical features are fever and skin rashes on the hands, feet, and oral cavity. Maculopapular rash and herpes appear on the hands, feet and buttocks, which mostly occur in preschool children, especially in the age group under 3 years old. This type of disease is distributed all over the world and occurs throughout the year in the tropics and subtropics. [0003] The main causes of hand, foot and mouth disease are ty...

AI Technical Summary

Problems solved by technology

The above-mentioned detection kits have the advantages of specificity, sensitivity, rapidity, and easy operation for detecting the corresponding enteroviruses, but none of the above-mentioned kits can realize the universal detection of enteroviruses and enteroviruses EV71, CoxA16, etc. in the same reaction tube. At the same time, a large number of experimental data show that the proportion of PCR inhibitors in stool, rectal swab, and throat swab samples is 1.8% to 3.4%, indicating that the internal control is important for monitoring the presence of PCR inhibitors. At present, the existing in vitro diagnostic kits for hand, foot and mouth disease have no internal control and have defects such as low specificity and sensitivity, and cumbersome detection steps, which need to be further improved.

Images