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 according 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 H1N1 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 diagnosis of influenza A virus (H1N1) for basic medical units.
白志军, 胡林, 李魁彪, 钟华燕, 陈艺韵, 鲁恩洁, 狄飚. 交叉引物恒温扩增法检测甲型H1N1流感病毒及临床应用[J]. 中国人兽共患病学报, 2015, 31(3): 208-211.
BAI Zhi-jun, HU Lin, LI Kui-biao, ZHONG Hua-yan, CHEN Yi-yun, LU En-jie, DI Biao. Establishment of cross priming amplification for influenza A virus (H1N1) and its clinical application. Chinese Journal of Zoonoses, 2015, 31(3): 208-211.
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