Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Acta Agriculturae Zhejiangensis》 2019-01
Add to Favorite Get Latest Update

Establishment of a multiplex PCR assay for three pathogenic bacteria in cows with recessive mastitis

WANG Yu;LI Bi;YANG Mingxian;ZUO Zhicai;College of Veterinary Medicine,Sichuan Agricultural University,Environmental Pollution and Animal Diseases of Sichuan Provincial Key Laboratory of Colleges and Universities,Sichuan Key Laboratory of Animal Diseases and Human Health;  
In order to establish a method for the rapid and simultaneous detection of three pathogenic bacteria of mastitis of Streptococcus agalactiae,Staphylococcus aureus and Pseudomonas aeruginosa,three pairs of specific primers were synthesized based on the Streptococcus agalactiae 16 S rRNA gene,Staphylococcus aureus NUC heat-resistance gene,Pseudomonas aeruginosa ETA gene sequence and references. By adjusting and optimizing the parameters of the PCR reaction,a multiplex PCR detection system was established,and the sensitivity test and specificity test were simultaneously performed. The results showed that the detection method can simultaneously amplify specific fragments of 270 bp of Streptococcus agalactiae,153 bp of Staphylococcus aureus and 375 bp of Pseudomonas aeruginosa and a generic 1 500 bp fragment of bacterial 16 S rRNA. For other six kinds of pathogenic bacteria,only 1 500 bp universal fragments could be amplified,which had strong specificity. Three specific primer amplification bands were 99% homologous to the Gen Bank and bacterial gene sequences. After culture for 24 h,the minimum sensitivity of the system to Streptococcus agalactiae,Staphylococcus aureus and Pseudomonas aeruginosa was 8 × 10~4,4 × 10~4 and 1. 5 × 10~5 CFU·m L~(-1),respectively. The detection method established in this study can complete the rapid detection of Streptococcus agalactiae,Staphylococcus aureus and Pseudomonas aeruginosa,and is of great significance for the differential diagnosis and rapid detection of subclinical mastitis in dairy cows.
【Fund】: 四川省科技厅重点研发项目(18ZDYE3215);; 优质肉牛高效安全生产技术研发与产业化示范;; 现代农业产业技术体系四川肉牛创新团队资助项目(035Z2036)
【CateGory Index】: S858.23
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
【Citations】
Chinese Journal Full-text Database 7 Hits
1 KAN Wei;WANG Hua;MA You-ji;ZHAO Xing-xu;ZHANG Yong;College of Veterinary Medicine,Gansu Agricultural University;Faculty of Animal Science and Technology,Gansu Agricultural University;;Construction of Eukaryotic Expression Vector of cfb Gene from Bovine Streptococcus agalactiaee[J];华北农学报;2014-05
2 LI Dong-liang;KOU Ya-nan;SONG Yue;WEI Zhan-yong;WANG Ya-bin;College of Animal Science and Veterinary Medicine,Henan Agricultural University;Key Laboratory for Animal-derived Food Safety of Henan Province;;Detection of Five Bacterial Species in Bovine Mastitis by PCR Based on 16S rDNA Sequence[J];河南农业科学;2014-06
3 GAO Gui-sheng;GAO Guang-ping;LI Zheng-ben;SHI Qiu-mei;ZHANG Yan-ying;SHAO Xin-hua;LIANG Yin-ju;Key Laboratory of Preventive Veterinary Medicine in Hebei Province /Animal Science School,Hebei Normal University of Science & Technology;Qian'an Jinfuzhi Dairy cattle farm;Hebei Xinhua Scientific Veterinary Medicine Co.,Ltd;;The Investigation of Main Pathogens Causing Dairy Cattle Recessive Mastitis in Eastern Hebei[J];Animal Husbandry and Feed Science;2013-04
4 HAN Qing-song,JIAN Yong-li,TU Yi-qiang,GAO Yong-an,SONG Xian-zhang,TU Guo-zhong,DUAN Long-chuan (Wenzhou Vocational College of Science and Technology,Wenzhou 325006, China);Research Advances on Pseudomonas aeruginosa[J];畜牧与饲料科学;2012-01
5 YE Fen,CAO Jie,WEI Xue-liang(College of Animal Science and Technology,Southwest University,Beibei,Chongqing 400716,China);Research Advance of Prevention and Treatment of Mastitis in Dairy Cattle[J];中国牛业科学;2007-05
6 QIN Fang-yun1,2, XIONG Yi1, SHENG Yu-ming2, BAI Yun2, QIN Lun2, ZHU Wei2,LI Hua-ming1, PAN Jie2, LONG Jian-ming2, CHEN Lei2,LIU Qi1(1.Guangxi General Veterinary Prevention and Quarantine Services, Nanning 530001, China; 2.College of Animal Science and Technology, Guangxi University, Nanning 530005, China);Establishment and preliminary application of multiplex PCR for rapid detection of Streptococcus suis serotype 2[J];畜牧与兽医;2007-09
7 MA Bao-chen~1,QIN Zhuo-ming~2,CAI Yu-mei~1,DONG Yu-lan~3,CHAI Tong-jie~(1*)(1.College of Animal Science and Technology,Shandong Agricultural University, Tai'an,271018,China;2.Shandong Academy of Agricultural Sciences,Ji'nan 250000,China;3.College of Veterinary Medicine,China Agricultural University,Beijing 100094,China);Development of Mutiplex PCR for Simultaneous Detection of Staphycococcus aureus,Streptococcus agalactiae,Streptococcus dysgalactiae and Yeasts in Bovine Mastitis Milk Samples[J];畜牧兽医学报;2006-11
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 FENG Jie;XIE Jian-yun;WEI Xiao-feng;HU Jian-hua;GAO Cheng;College of Veterinary Medicine,Yangzhou University;Shanghai Laboratory Animal Research Center,Shanghai Quality Monitoring Center of Laboratory Animals;;Molecular Subtyping Study on Pseudomonas aeruginosa in Rats and Mice with Pulsed Field Gel Electrophoresis[J];实验动物与比较医学;2016-06
2 HONG Ying;XIAO Zhen;HAO Jun-Hu;JIANG Lu-Yan;WANG Yi-Qian;WU Fu-Ping;SHAO Jing-Dong;JIANG Yuan;FENG Li;HUANG Ming;ZHU Chang-Qing;Jiangsu Entry-Exit Inspection and Quarantine Bureau;Ningxia Entry-Exit Inspection and Quarantine Bureau;College of Food Science and Technology,Nanjing Agricultural University;;Comparison of different methods for DNA extraction of Streptococcus agalactiae in milk[J];食品安全质量检测学报;2015-12
3 LIU Xi;AO Qiu-wei;TAN Yun;ZHOU Yu;GAN Xi;ZHU Jia-jie;Guangxi Academy of Fishery Science/Guangxi Key Lab of Aquatic Genetic Breeding and Healthy Aquaculture;College of Animal Science and Technology,Guangxi University;;Dynamic distribution of pathogenic bacteria in GIFT infected artificially with Streptococcus agalactiae[J];南方农业学报;2015-09
4 Liu Longhai;Li Xinpu;Li Hongsheng;Key Laboratory of New Animal Drug Project of Gansu Province,Key Laboratory of Veterinary Pharmaceutical Discovery ,Ministry of Agriculture,Lanzhou Institute of Animal Husbandry and Pharmaceutical Sciences ,Chinese Academy of Agricultural Sciences;;Progress in the Key Technologies for Controlling Bovine Mastitis[J];中国草食动物科学;2015-05
5 ZHANG Xiang-zhai;ZHANG Yan-ying;LI Yun-yu;LI Pei-guo;JIA Qing-hui;ZHANG Jian-wen;Key Laboratory of Preventive Veterinary Medicine in Hebei Province,Hebei Normal University of Science and Technology;;Isolation,Identification and Drug Susceptibility Test of Pseudomonas aeruginosain Chickens[J];中国畜牧兽医;2015-06
6 HE Miao;ZHANG Guo-li;CHEN Ping;College of Food Science and Engineering,Jilin Agricultural University;;Study on the expression,purification and immune efficacy of fusion protein HSP65-PEAⅠ[J];中国实验诊断学;2015-05
7 LAI Jin-lun;LIU Yu-hui;KOU Ming-ming;LIU Jing;HU Chang-min;GUO Ai-zhen;The Faculty of Veterinary Medicine, Huazhong Agricultural University;Huazhong Agricultural University, State Key Laboratory of Agricultural Microbiology;;Isolation, Identification and Drug Resistance of Staphylococcus Aureus from Milk[J];中国奶牛;2015-09
8 KONG Na;Department of Pharmaceutical Science and Technology,Heze University;;Curative Effect of Combination of Traditional Chinese and Western Medicines Treatment on Cows Mastitis[J];河南农业科学;2015-03
9 JIANG Xiao-dong;SHAO Ming-xu;LIU Li-ping;WANG Chuan-wen;YU Cui-lian;CHI Shan-shan;ZHU Rui-liang;Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention/College of Animal Science and Technology,Shandong Agricultural University;;Isolation and phylogenetic analysis of sheep Pseudomonas aeruginosa from partial areas in Shandong[J];中国兽医学报;2015-03
10 LI Yueying;ZHU Song;JU Huidong;LIU Wenqing;ZHAO Baohua;PEI Yantao;College of Life Science,Hebei Normal University;;Cloning Expression and Immunogenicity Study of ISP Protein of Streptococcus dysgalactiae[J];河北师范大学学报(自然科学版);2015-02
【Secondary Citations】
Chinese Journal Full-text Database 10 Hits
1 LI Su-ying(Biotechnology Department of Ulanqab Vocational College of Inner Mongolia,Ulanqab 012000,China);Curative Effect of Integrative Medicine on Cow Clinical Mastitis[J];河南农业科学;2013-05
2 WANG Dong1,SUN Ying-feng1,CHI Jing-jing1,YU Hua-fu2,WANG Ying-zhen1 (1. Tianjin Institute of Animal Science and Veterinary Medicine,Tianjin 300112,China;2. Seventh Farm of Dairy Cattle in Jialihe Animal Husbandry Limitted Company,Tianjin 301508,China);Screening of Sensitized Drugs in Mycotic Mastitis of Dairy Cattle[J];天津农业科学;2012-03
3 ZHANG Shan-rui1.2,WANG Chang-fa1,GAO Yun-dong1,YANG Shao-hua1, ZHAO Hong-kun1.2,ZHONG Ji-feng1(1.Cow Research Center of Agricultural Science Academy of Shandong Province,Jinan,Shandong 250100,China;2.College of Veterinary Medicine,Shandong Agricultural University,Taian,Shandong 271018,China);Identification of Major Pathogens in Bovine Mastitis by Multiplex PCR[J];家畜生态学报;2007-04
4 YU Dan-feng,KONG Fan-zhi,ZHU Wan-ping,LI Jing.Zhejiang Tongde Hospital,Hangzhou 310012,Zhejiang,China;Experimental study on anti-pseudomonas aeruginosa infection in trachea by astragalus polysaccharides(黄芪多糖)[J];中国中西医结合急救杂志;2007-02
5 Ai Qijun1 Yu Qinghua1,2 Zhang Hongxing1 Zhang Xinru1 Liu Yiqian1 Piao Jingang1(1Department of Food Science,Beijing University of Agriculture,Beijing 102206 2Food Science and Engineering Department,Inner Mongolia Agriculture University,Huhhot 010018);Detection of the Gene Encoding Exotoxin A of Pseudomonas Aeruginosa by PCR Technique[J];中国食品学报;2006-06
6 XIE Zhi-qin,XIE Zhi-xun~*,TANG Xiao-fei,LIU Jia-bo,PANG Yao-shan,DENG Xian-wen,ZHU Xian-long (Guangxi Veterinary Research Institute,Nanning 530001,China);Development of a rapid and sensitive multiplex PCR for detection of three major pathogens in bovine mastitis[J];畜牧与兽医;2006-07
7 WU Run,HAO Bao-qing,NONG Xiang,YAN Dan-hong(College of Life Science and Technology,Southwest Nationalities University,Chengdu,Sichuan 610041,China);Identification of Major Pathogens in Bovine Subclinic Mastitis by PCR[J];中国牛业科学;2006-02
8 ZHAO Hua-mei,PAN Xiu-zhen,WANG Chang-jun,GUO Heng-bin,LI Xian-fu,TANG Jia-qi(Institute of Medical Molecular Biology,Nanjing Normal University,Nanjing 210097,China);Cloning,expression and identification of gene encoding the glutamate dehydrogenase of Streptococcus suis serotype 2[J];中国人兽共患病学报;2006-01
9 LU Cheng-ping School of Veterinary Medicine, Nanjing Agricultural University,Nanjing 210095, China;Swine Streptococcosis and Streptococcus Suis Type 2[J];科技导报;2005-09
10 ZHOU Xue-zhang, DENG Xu-ming (College of Animal Science and Technology, The Quartermaster University of PLA ,Changchun Jilin 130062 China);Studying of the disease of pseudomonas aeruginosa in chickens[J];中兽医医药杂志;2004-03
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved