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Open Access Highly Accessed Research article

Development and implementation of the quality control panel of RT-PCR and real-time RT-PCR for avian influenza A (H5N1) surveillance network in mainland China

Rongbao Gao1, Yan Gao1, Leying Wen1, Ming Shao2, Shumei Zou1, Changgui Li2, Lei Yang1, Xiyan Li1, Wei Wang1 and Yuelong Shu1*

Author Affiliations

1 Dept. of Influenza, Chinese National Influenza Center, State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, Chinese center for disease control and prevention (China CDC), Beijing 100052, PR China

2 The 3rd Division of Viral Vaccine, the National Institute for the Control of Pharmaceutical and Biological Products, Beijing, 100050, PR China

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BMC Infectious Diseases 2011, 11:67  doi:10.1186/1471-2334-11-67

Published: 16 March 2011

Abstract

Background

Reverse transcription PCR (RT-PCR) and real time RT-PCR (rRT-PCR) have been indispensable methods for influenza surveillance, especially for determination of avian influenza. The movement of testing beyond reference lab introduced the need of quality control, including the implementation of an evaluation system for validating personal training and sample proficiency testing.

Methods

We developed a panel with lysates of seasonal influenza virus (H1N1, H3N2 and B), serials of diluted H5N1 virus lysates, and in-vitro transcribed H5 hemaglutinin (HA) and an artificial gene RNAs for RT-PCR and rRT-PCR quality control assessment. The validations of stability and reproducibility were performed on the panel. Additionally, the panel was implemented to assess the detection capability of Chinese human avian influenza networks.

Results

The panel has relatively high stability and good reproducibility demonstrated by kappa's tests. In the implementation of panel on Chinese human avian influenza networks, the results suggested that there were a relatively low number of discrepancies for both concise and reproducibility in Chinese avian influenza virus net works.

Conclusions

A quality control panel of RT-PCR and real-time RT-PCR for avian influenza A (H5N1) surveillance network was developed. An availably statistical data, which are used to assess the detection capability of networks on avian influenza virus (H5N1), can be obtained relatively easily through implementation of the panel on networks.