SYBR Green I Real-time PCR method for quickly detecting and indentifying Hantaan virus infection

Abstract

The invention discloses an SYBR Green I Real-time PCR (Real-time fluorescent quantitative polymerase chain reaction) method for quickly detecting and indentifying Hantaan virus infection. The method comprises the following steps: designing a pair of specific primers mainly according to a conserved sequence of an S gene in a GenBank database; using a positive plasmid subjected to 10-time gradient dilution as a standard product; optimizing a reaction condition; establishing the SYBR Green I Real-time PCR method used for detecting a Vero E6 cell, a mouse and a clinical patient, which are infected by Hantaan virus nucleic acids according to qRT-PCR specificity amplification, wherein a copy number of the detection sensitivity can reach 101, and sensitivity, repeatability and specificity of the Hantaan virus nucleic acids are detected. Compared with a conventional Hantaan virus detection method, the method has the advantages that types of detecting samples are rich, the application range is wide, the contrast is strong, operation steps are relatively few, convenience and quickness are realized, the sensitivity and the repeatability are high, the specificity is good, and a linear relationship is good, and can provide a strong experimental basis on clinical and laboratory detection of hemorrhagic fever with renal syndrome as well as studying of epidemiology.

Description

technical field [0001] The invention belongs to the field of virus detection and relates to related fields such as molecular biology, virology, immunology and immune application. The invention specifically relates to a SYBR GreenⅠReal-time PCR method for rapid detection and identification of Hantaan virus infection. The samples for detection and identification of Hantaan virus nucleic acid include infection of Vero E6 cells, infection of experimental mice and clinical patients. It includes designing a pair of primers for specifically amplifying Hantaan virus, preparing standards, researching and optimizing experimental conditions, and performing qRT-PCR specificity on total RNA extracted from infected Vero E6 cells, infected experimental mice and clinical patient serum Amplification detection. Background technique [0002] Research status of diagnosis and detection of hemorrhagic fever with renal syndrome: [0003] Hantaan virus belongs to the genus Hantavirus in the Buny...

AI Technical Summary

Problems solved by technology

However, the classic separation and identification method is time-consuming, and the IFA method is not suitable for rapid on-site detection. The detection of HTNV antibody is only an indirect indicator of one side, and there are problems such as insufficient specificity and sensitivity, and inaccurate quantification.

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