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

Abstract

The invention discloses a SYBR GreenⅠReal-time PCR method for rapid detection and identification of Hantaan virus infection. The method mainly designs a pair of specific primers based on the sequence of the conserved region of the S gene in the GenBank database. The gradient diluted positive plasmid was used as the standard, the reaction conditions were optimized, and the specific amplification by qRT-PCR was used to establish the SYBR Green Ⅰ Real-time PCR method for the detection of Hantaan virus nucleic acid in infected Vero E6 cells, infected mice and clinical patients, and the detection sensitivity A copy number of 101 was reached and tested for sensitivity, reproducibility, and specificity. Compared with conventional Hantaan virus detection and identification methods, this method has the advantages of rich types of detection specimens, wide application range, strong contrast, few operation steps, convenient and fast, high sensitivity, high repeatability and specificity, and has good The advantage of a linear relationship. It provides a strong experimental basis for the clinical, laboratory detection and epidemiological research of hemorrhagic fever with renal syndrome.

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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