Establishment of cross priming amplification for influenza A virus (H1N1) and its clinical application
10.3969/cjz.j.issn.1002-2694.2015.03.004
- VernacularTitle:交叉引物恒温扩增法检测甲型 H1N1流感病毒及临床应用
- Author:
Zhijun BAI
;
Lin HU
;
Kuibiao LI
;
Huayan ZHONG
;
Yiyun CHEN
;
Enjie LU
;
Biao DI
- Publication Type:Journal Article
- Keywords:
influenza A virus(H1N1);
cross priming amplification (CPA);
clinical application
- From:
Chinese Journal of Zoonoses
2015;(3):208-211,215
- CountryChina
- Language:Chinese
-
Abstract:
In this study ,we established Cross Priming Amplification (CPA ) technology for detection of influenza A virus (H1N1) approach ,and evaluated the method through clinical specimens .A set of specific primers were designed for CPA ac‐cording to the conservative gene sequences ,designed and realized in the same temperature reverse transcription of RNA and DNA amplification . The amplification products can be totally enclosed nucleic acid detection device for testing . Fourteen healthy pharyngeal swab specimens ,seven other respiratory viruses ,and six arboviruses strains were used as the controls .We used a method that application of gradient dilution to the H 1N1 virus strain as the control to test the sensitivity of the CPA .We also used 102 clinical pharyngeal swab specimens of H1N1 patients for detection object to evaluate the feasibility of CPA clinical detection .Results showed that the CPA reaction did not appear cross reaction on health cases samples and other viruses .The sensitivity of the CPA was approximately 10 copies/uL in the established method that exactly titer H1N1 virus strain gradient dilution test .As to the positive results among the clinical pharyngeal swab samples collected from patients at different stages after onset ,the CPA had the highest positive detection rate during the first three days after onset (100% ) .While the detection rate from day 4 to day 6 after onset was 79 .31% .After 7 days ,the detection rate was 9 .09% .The established CPA assay was a highly sensitive ,specific and reproducible approach for rapid detection of H1N1 virus ,which is conducive to the early diagno‐sis of influenza A virus (H1N1) for basic medical units .
