H3亚型禽流感病毒巢式PCR检测方法的建立

Abstract

Abstract: In order to establish a method to detect H3 subtype avian influenza virus (AIV), two pairs of specific primers were designed and selected according to the conserved sequences of the hemagglutinin (HA) genes of H3 subtype AIV, the reaction conditions were optimized to establish the nested PCR.The specificity and sensitivity of assays were evaluated, and 96 clinical samples were detected by nested PCR. The results showed that the nested PCR was only specific to H3 subtype AIV, without amplification of other viruses; the detection limit of nested PCR was 1×10³ copies/μL which was 100 fold higher than the routine PCR; the detection of clinical samples were consistent with the viral isolation completely. This study provided a accurate and effective method for the detection of H3 subtype AIV.

Key words: H3 subtype avian influenza virus (AIV); nested PCR; detection method

CLC Number:

LIU Ting-ting, XIE Zhi-xun, SONG De-gui, LUO Si-si, XIE Li-ji, LI Meng, XIE Zhi-qin, DENG Xian-wen. Establishment of Nested PCR for Diagnosis of H3 Subtype Avian Influenza Virus[J]. , 2014, 41(11): 85-89.

References

1 王莹, 谢芝勋,刘加波,等. 禽呼肠孤病毒四种核酸检测方法的比较[A].中国畜牧兽医学会禽病学分会第十五次学术研讨会论文集[C].2010.
2 甘孟侯. 禽流感[M]. 北京:北京农业大学出版社,2002.
3 杨素,廖秀云,廖明,等.禽流感病毒通用型及H5亚型套式荧光RT-PCR检测方法研究[J].中国畜牧兽医,2012,39(3):165～170.
4 周辰瑜,谢芝勋,彭宜,等. H6亚型禽流感病毒巢式RT-PCR检测方法的建立[J].畜牧与兽医,2012,44(11):12～16.
5 唐熠,谢芝勋,刘加波,等.Ⅰ群禽腺病毒四种核酸检测方法的比较[A].中国畜牧兽医学会禽病学分会第十五次学术研讨会论文集[C]. 2010.
6 郭捷, 谢芝勋, 彭宜, 等. H1和H3亚型禽流感病毒二重PCR检测方法的建立[J].动物医学进展,2012,33(12):31～34.
7 郭捷, 谢芝勋, 罗思思, 等. 巢式PCR检测H1亚型禽流感病毒方法的建立[J].中国畜牧兽医,2013,40(9):23～26.
8 韩雪清, 林祥梅, 侯义宏, 等. 禽流感病毒H1、H3、H5、H2亚型多重RT-PCR检测方法的初步研究[J].中国人兽共患病学报,2007,23(10):993～996.
9 Alexander D J. A review of avian influenza in different bird species[J]. Vet Microbiol, 2000, 74(1～2):3～13.
10 Campitelli L, Fabiani C, Puzelli S, et al. H3N2 influenza viruses from domestic chickens in Italy: An increasing role for chickens in the ecology of influenza[J]. Gen Virol, 2002, 83:413～420.
11 Duan L, Campitelli L, Fan X H, et al. Characterization of low-pathogenic H5 subtype influenza viruses from Eurasia: Implications for the origin of highly pathogenic H5N1 viruses[J]. J Virol, 2007, 81(14):7529～7539.
12 Peng Y, Xie Z, Liu J, et al. Visual detection of H3 subtype avian influenza viruses by reverse transcription loop-mediated isothermal amplification assay[J]. Virology Journal, 2011, 8:337.
13 Peng Y, Xie Z, Liu J, et al. Epidemiological surveillance of low pathogenic avian influenza virus (LPAIV) from poultry in Guangxi province, Southern China[J]. PLoS One, 2013, 8(10):e77132.
14 Song M S, Oh T K, Moon H J, et al. Ecology of H3 avian influenza viruses in Korea and assessment of their pathogenic potentials[J]. J Gen Virol, 2008, 89:949～957.
15 Webster R G, Bean W J, Gorman O T, et al. Evolution and ecology of influenza A viruses[J]. Microbiol Rev, 1992, 56:152～179.
16 Xie Z, Xie L, Zhou C, et al. Complete genome sequence analysis of an H6N1 avian influenza virus isolated from Guangxi pockmark ducks[J]. J Virol, 2012, 86:13868～13869.

[1]	CHEN Jimin, LI Huihuang, LAI Lianjie, YANG Yinhua, YANG Yue, LIN Ruiyi. Advance on Detection Methods and Vaccines of Duck Tembusu Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1329-1339.
[2]	WANG Qianqian, YU Yongle, LI Yuehua, DONG Yaqin, NI Bo, LIU Fuxiao. Advance in Detection Methods and Vaccines of Senecavirus A [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2336-2346.
[3]	GUO Luanying, LI Jialei, JIN Qinqin, MA Jun, LIU Quan. Research Progress on Bovine Hepacivirus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1994-2000.
[4]	WU Shuaibin, CUI Zehua, LIAO Xiaoping, LIU Yahong, SUN Jian. Research Progress on the Detection Methods of Carbapenemase-producing Enterobacteriaceae [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 1072-1083.
[5]	WANG Yong, HAN Yuqi, LU Minghua. Research Progress on Detection Methods of Drug Residues of β-receptor Agonist [J]. , 2019, 46(8): 2478-2487.
[6]	LIU Dingxi, YANG Jiecheng, ZHOU Hongchao, GUO Kangkang. Research Advance on Detection Methods of Antibacterial Residues in Animal Food [J]. , 2019, 46(7): 2183-2192.
[7]	WANG Yong, GONG Yong, LU Ming-hua. Research Progress on Detection Methods of Fluoroquinones Residues [J]. , 2017, 44(8): 2509-2516.
[8]	CHEN Shao-pin, LIN Han-qing, WEN Gui-lan, ZHANG Sheng-bo, LI Chang-hong, XU Li, GONG Xin-yong, WANG De-sheng, WEN Ming, ZHOU Bi-jun, CHENG Zhen-tao. Nested PCR Amplification and Bioinformatics Analysis of LSm1 Gene from Marc145 Cells [J]. , 2017, 44(6): 1604-1612.
[9]	HAN Xiao-ya, CHEN Dong-mei, WANG Yu-lian, TAO Yan-fei, DONG Guo-liang, PENG DA-peng, YUAN Zong-hui. Research Progress on Antibodies Rapid Detection Technology for Veterinary Synthetic Antibacterial Agents [J]. , 2017, 44(6): 1869-1876.
[10]	FENG Bin, XIE Zhi-xun, ZHANG Yan-fang, HUANG Jiao-ling, WANG Sheng, FAN Qing, HUANG Li, XIE Li-ji, ZENG Ting-ting, LUO Si-si, DENG Xian-wen, XIE Zhi-qin, LIU Jia-bo, SONG Zhong-bao, LIN Er-ke. Development and Preliminary Application of a Semi-nested PCR Assay for Detection of Chicken Parvovirus [J]. , 2017, 44(5): 1295-1301.
[11]	YAO Huai-bing, ZHAO Yi, LIU Hong, ZHANG Yi, REN Fang, HUANG Jiong. Development and Application of Two-temperature RT-PCR for Detectionof Type O Foot and Mouth Disease Virus [J]. , 2017, 44(2): 311-318.
[12]	XU Ming-yu, SONG Shu-ting, ZHANG Ying, ZHANG Hui-tu. Improved Biotin Detection Method of Enduracidinmold [J]. , 2017, 44(2): 384-390.
[13]	FENG Chun-yan, DU Fang-yuan, LIU Dan-dan, Pershin Andrey, WANG Cai-xia, ZHANG Yong-ning, WU Shao-qiang, LIN Xiang-mei. Development of Virus-like Particles Containing African Swine Fever Virus Nucleic Acid Sequence and Its Application in Detection Method [J]. , 2017, 44(11): 3287-3293.
[14]	WANG Ming-ming, XU Na, TANG Yun, HANG Ye, LIU Zhen-jiang, DU Dao-lin. Advances on Detection Methods for Residues of Nitrofurans in Food [J]. , 2016, 43(8): 2202-2207.
[15]	ZHU Yu-fei, XIE Shu-yu, CHEN Dong-mei, WANG Yu-lian, PAN Yuan-hu, PENG Da-peng, TAO Yan-fei, YUAN Zong-hui. Research Progress on Detection Methods of Feed Additives [J]. , 2016, 43(7): 1785-1791.

Establishment of Nested PCR for Diagnosis of H3 Subtype Avian Influenza Virus

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments