A rapid and sensitive loop-mediated isothermal amplification procedure (LAMP) for Mycoplasma hyopneumoniae detection based on the p36 gene

文献类型： 外文期刊

第一作者： Liu, M. J.

作者： Liu, M. J.;Du, G. M.;Bai, F. F.;Wu, Y. Z.;Xiong, Q. Y.;Feng, Z. X.;Li, B.;Shao, G. Q.;Du, G. M.;Liu, M. J.;Shao, G. Q.

作者机构：

关键词： p36 gene;Loop-mediated isothermal amplification;Mycoplasma hyopneumoniae

期刊名称：GENETICS AND MOLECULAR RESEARCH （ 影响因子：0.764； 五年影响因子：0.912 ）

ISSN： 1676-5680

年卷期： 2015 年 14 卷 2 期

页码：

收录情况： SCI

摘要： The aim of this study was to establish a method for sensitive and rapid diagnosis of Mycoplasma hyopneumoniae in clinical specimens. To this effect, we employed three sets of primers specifically designed for amplification of nucleic acids under isothermal conditions. After optimization of reaction conditions, M. hyopneumoniae could be successfully detected at 63 degrees C in 45 min through use of the loop-mediated isothermal amplification (LAMP) assay. A positive reaction was identified visually as white precipitate and confirmed by gel electrophoresis. The detection limit for this assay was 10 copies/mu L, as observed by electrophoretic analysis. The accuracy of the LAMP reaction was confirmed by restriction endonuclease digestion as well as by direct sequencing of the amplified product. This method can specifically detect M. hyopneumoniae; other species with high homology and other bacterial and virus strains gave negative results. To test the utility of this procedure, the LAMP assay was applied to 40 clinical samples collected from swine lung tissues experimentally challenged with M. hyopneumoniae isolates, and compared to the results from a real-time polymerase chain reaction (PCR) assay. A concordance of 100% was observed between the two assays. In conclusion, the results from our study demonstrated that the LAMP assay provided a rapid reaction and was inexpensive to perform, with no need of complex instruments or systems such as Geneamp PCR. The LAMP assay may therefore be applied in routine diagnosis in the clinical laboratory and for in-field detection of M. hyopneumoniae infection.

分类号：

  • 相关文献

[1]Effects of Mycoplasma hyopneumoniae on porcine nasal cavity dendritic cells. Shen, Yumeng,Hu, Weiwei,Yang, Qian,Wei, Yanna,Feng, Zhixin.

[2]In vitro protective efficacy of Lithium chloride against Mycoplasma hyopneumoniae infection. Ishag, Hassan Z. A.,Wu, Yu-zi,Liu, Mao-jun,Xiong, Qi-yan,Feng, Zhi-xin,Yang, Ruo-song,Shao, Guo-qing,Ishag, Hassan Z. A..

[3]Transcription analysis on response of porcine alveolar macrophages to co-infection of the highly pathogenic porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae. Li, Bin,Du, Luping,Xu, Xiangwei,Sun, Bing,Yu, Zhengyu,Feng, Zhixin,Liu, Maojun,Wei, Yanna,Wang, Haiyan,Shao, Guoqing,He, Kongwang,Li, Bin,Yu, Zhengyu,Feng, Zhixin,Liu, Maojun,Wei, Yanna,Wang, Haiyan,Shao, Guoqing,He, Kongwang.

[4]Loop-mediated isothermal amplification for rapid and convenient detection of Mycoplasma hyopneumoniae. Li, Jiahe,Jiang, Fei,Yang, Sheng,Guo, Panpan,Wu, Wenxue,Minion, F. Chris,Petersen, Andrew C.,Li, Jinxiang.

[5]The effects of Mycoplasma hyopneumoniae on porcine circovirus type 2 replication in vitro PK-15 cells. Wang, Haiyan,Feng, Zhixin,Wu, Yuzi,Wei, Yanna,Gan, Yuan,Hua, Lizhong,Li, Bin,Wang, Xiaomin,Liu, Maojun,Xiong, Qiyan,Shao, Guoqing,Wang, Haiyan,Feng, Zhixin,Shao, Guoqing.

[6]Protective efficacy of a live attenuated Mycoplasma hyopneumoniae vaccine with an ISCOM-matrix adjuvant in pigs. Xiong, Qiyan,Wei, Yanna,Feng, Zhixin,Gan, Yuan,Liu, Zhanjun,Liu, Maojun,Bai, Fangfang,Shao, Guoqing.

[7]CYP1A1 mediates the suppression of major inflammatory cytokines in pulmonary alveolar macrophage (PAM) cell lines caused by Mycoplasma hyponeumoniae. Fang, Xiaomin,Zhao, Weimin,Xu, Jie,Tu, Feng,Wang, Xuemin,Li, Bixia,Fu, Yanfeng,Ren, Shouwen.

[8]Use of serological and mucosal immune responses to Mycoplasma hyopneumoniae antigens P97R1, P46 and P36 in the diagnosis of infection. Feng, Zhi-Xin,Bai, Yun,Yao, Jing-Ting,Gan, Yuan,Wang, Hai-Yan,Wei, Yan-Na,Liu, Mao-Jun,Xiong, Qi-Yan,Bai, Fang-Fang,Li, Bin,Wu, Xu-Su,Shao, Guo-Qing,Feng, Zhi-Xin,Bai, Yun,Liu, Mao-Jun,Shao, Guo-Qing,Pharr, G. Todd,Wan, Xiu-Feng,Xiao, Shao-Bo,Chi, Ling-Zhi.

[9]Development of a blocking ELISA for detection of Mycoplasma hyopneumoniae infection based on a monoclonal antibody against protein P65. Liu, Maojun,Zhang, Yue,Wu, Yuzi,Wang, Haiyan,Li, Bin,Bai, Yun,Feng, Zhixin,Xiong, Qiyan,Bai, Fangfang,Shao, Guoqing,Liu, Maojun,Du, Gaimei,Browning, Glenn F.,Shao, Guoqing.

[10]Comparative analysis of mucosal immunity to Mycoplasma hyopneumoniae in Jiangquhai porcine lean strain and DLY piglets. Hua, L. Z.,Wu, Y. Z.,Bai, F. F.,William, K. K.,Feng, Z. X.,Liu, M. J.,Yao, J. T.,Zhang, X.,Shao, G. Q.. 2014

[11]A replicating plasmid-based vector for GFP expression in Mycoplasma hyopneumoniae. Ishag, H. Z. A.,Liu, M. J.,Yang, R. S.,Xiong, Q. Y.,Feng, Z. X.,Shao, G. Q.,Ishag, H. Z. A.. 2016

[12]Mycoplasma hyopneumoniae induces pro-inflammatory cytokine and nitric oxide production through NF kappa B and MAPK pathways in RAW264.7 cells. Hwang, Mi-Hyun,Damte, Dereje,Lee, Joong-Su,Gebru, Elias,Chang, Zhi-Qiang,Rhee, Man-Hee,Park, Seung-Chun,Cheng, Henrique,Jung, Byeong-Yeal,Chang, Zhi-Qiang. 2011

[13]Development and validation of an attenuated Mycoplasma hyopneumoniae aerosol vaccine. Feng, Zhi-Xin,Wei, Yan-Na,Li, Gui-Lan,Lu, Xiao-Ming,Wang, Zhan-Wei,Kong, Meng,Gan, Yuan,Bai, Fang-Fang,Liu, Mao-Jun,Xiong, Qi-Yan,Wu, Xu-Su,Shao, Guo-Qing,Wan, Xiu-Feng,Pharr, G. Todd. 2013

[14]Cholesterol exacerbates Mycoplasma hyopneumoniae-induced apoptosis via stimulating proliferation and adhesion to porcine alveolar macrophages. Liu, Maojun,Hu, Yun,Liu, Jie,Jia, Yimin,Zhao, Ruqian,Liu, Maojun,Liu, Beibei,Shao, Guoqing,Jia, Yimin,Zhao, Ruqian,Du, Gaimei,Minion, F. Chris,Shao, Guoqing. 2017

[15]Comparative genomic analyses of Mycoplasma hyopneumoniae pathogenic 168 strain and its high-passaged attenuated strain. Liu, Wei,Xiao, Shaobo,Li, Mao,Guo, Shaohua,Li, Sha,Luo, Rui,Chen, Huanchun,Fang, Liurong,Feng, Zhixin,Li, Bin,Shao, Guoqing,Zhou, Zhemin. 2013

[16]Apoptosis induced by lipid-associated membrane proteins from Mycoplasma hyopneumoniae in a porcine lung epithelial cell line with the involvement of caspase 3 and the MAPK pathway. Ni, B.,Bai, F. F.,Wei, Y.,Liu, M. J.,Feng, Z. X.,Xiong, Q. Y.,Hua, L. Z.,Shao, G. Q.. 2015

[17]Vaccination inhibits TLR2 transcription via suppression of GR nuclear translocation and binding to TLR2 promoter in porcine lung infected with Mycoplasma hyopneumoniae. Sun, Zhiyuan,Liu, Maojun,Zou, Huafeng,Li, Xian,Zhao, Ruqian,Liu, Maojun,Shao, Guoqing. 2013

[18]Effect of different adjuvant formulations on the immunogenicity and protective effect of a live Mycoplasma hyopneumoniae vaccine after intramuscular inoculation. Wang, Chunlai,Xie, Fang,Xiong, Qiyan,Wei, Yanna,Xie, Haidong,Feng, Zhixin,Gan, Yuan,Liu, Maojun,Bai, Fangfang,Shao, Guoqing.

[19]Immune Responses to the Attenuated Mycoplasma hyopneumoniae 168 Strain Vaccine by Intrapulmonic Immunization in Piglets. Feng Zhi-xin,Shao Guo-qing,Liu Mao-jun,Wu Xu-su,Zhou Yong-qi,Gan Yuan. 2010

[20]Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene. Zhuang, L.,Gong, J.,Wang, C.,Gong, J.,Zhu, C.,Yu, Y.,Dou, X.,Liu, X.,Xu, B.,Li, Q.,Zhuang, L.,Wang, C..

作者其他论文 更多>>

微信小程序