Real-time fluorescence RT-PCR method for quantitatively detecting human-infected avian influenza virus (A/H7N9)

Abstract

The invention belongs to the field of virus detection, and particularly relates to a real-time fluorescence RT-PCR method for quantitatively detecting a human-infected avian influenza virus (A / H7N9). According to comparative analysis with an existing human-infected H7N9 virus in a Genbank database, it is shown that a synthesized and designed primer and probe are completely conservative to the epidemic H7N9 virus. According to the method, a higher detection sensitivity is achieved on the condition that five genes are copied in each reaction, a cross reaction on 17 common respiratory tract infection viruses does not occur, and the H7N9 can be specifically detected; contrast detection between the method and a national CDC detection method is conducted on 80 clinical specimens, double-check is conducted on 13 inconsistent specimens by adopting rRT-PCR recommended by WTO for detecting an influenza A virus, detection results are all positive, and it is shown that the detection results are accurate and reliable. The method can conduct quantitative analysis on the copy number of the H7N9 virus in the clinical specimens, the specificity, the sensitivity and the accuracy are achieved, the method is suitable for large-scale detection of the clinical specimens, and an important basis is provided for optimizing clinical treatment strategies.

Description

technical field [0001] The invention belongs to the field of virus detection and relates to a detection method for human-infected avian influenza virus (A / H7N9), in particular to a real-time fluorescent RT-PCR method for quantitative detection of human-infected avian influenza virus (A / H7N9). Background technique [0002] According to reports, human infection with avian influenza virus (avian influenza virus, AIV) subtype H7N9 was first discovered in a certain area of ​​China in March 2013. As of October 24, more than 100 cases of human infection with avian influenza virus (A / H7N9) have been monitored, of which more than 40 cases died. The sudden outbreak has drawn great attention from the WHO and governments around the world. It was previously reported that the AIV subtypes directly transmitted from poultry to humans mainly include H5N1, H7N2, H7N3, H7N7, H9N2 and H10N7; Except for the 60% mortality rate, several other subtypes mainly cause symptoms of conjunctivitis or u...

AI Technical Summary

Problems solved by technology

However, according to the study by Hu et al., it was found that the H7N9 virus may undergo drug-resistant mutations during Tamiflu treatment, resulting in persistent virus positive. Therefore, it is necessary to pay close attention to the trend of viral load changes in patients during antiviral treatment in order to judge the antiviral effect in a timely manner. Treatment effect, providing basis for clinical optimal treatment plan

Although the National Influenza Center and foreign research group VMCorman and others have established real-time fluorescent RT-PCR methods to detect H7 and N9 genes respectively, they have not conducted quantitative research on the viral load in clinical specimens; in order to deal with sudden outbreaks, relevant prevention and control The department urgently needs to establish a method for rapid identification and detection of the new type of avian influenza A / H7N9, especially the urgent need to establish a specific, sensitive and reliable H7N9 viral load monitoring method suitable for large-scale clinical detection

Images