{"title":"环介导等温扩增法快速检测牛乳样品中的MRSA","authors":"G. Sathish, E. Hemakumar, K. Divya","doi":"10.9734/BMRJ/2016/28037","DOIUrl":null,"url":null,"abstract":"Aim: To develop a simple and rapid detection method for diagnosis of MRSA in bovine milk samples suspected for mastitis. Methodology: The laboratory sensitivity and specificity of LAMP assay was carried out using available laboratory strains. Milk samples were collected from Thiruvallur and Kanchipuram districts of Tamilnadu, India. DNA was directly isolated from the milk samples and mecA gene was screened by both PCR and LAMP methods. Results: The LAMP assay successfully amplified the mecA gene under isothermal conditions at 64°C within 60 min. LAMP assay was able to detect 10 pg of DNA and did not amplify mecA gene from bacterial DNA of other species. The screening of milk samples for MRSA showed 47 Positive out of 77 Samples by PCR and 43 positive out of 77 Samples by LAMP. Conclusion: Application of LAMP assay enabled rapid and easy detection of MRSA in milk samples suspected for bovine mastitis. Short Research Article Sathish et al.; BMRJ, 17(1): 1-5, 2016; Article no.BMRJ.28037 2","PeriodicalId":9269,"journal":{"name":"British microbiology research journal","volume":"18 1","pages":"1-5"},"PeriodicalIF":0.0000,"publicationDate":"2016-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Rapid Detection of MRSA by Loop Mediated Isothermal Amplification in Bovine Milk Samples\",\"authors\":\"G. Sathish, E. Hemakumar, K. Divya\",\"doi\":\"10.9734/BMRJ/2016/28037\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Aim: To develop a simple and rapid detection method for diagnosis of MRSA in bovine milk samples suspected for mastitis. Methodology: The laboratory sensitivity and specificity of LAMP assay was carried out using available laboratory strains. Milk samples were collected from Thiruvallur and Kanchipuram districts of Tamilnadu, India. DNA was directly isolated from the milk samples and mecA gene was screened by both PCR and LAMP methods. Results: The LAMP assay successfully amplified the mecA gene under isothermal conditions at 64°C within 60 min. LAMP assay was able to detect 10 pg of DNA and did not amplify mecA gene from bacterial DNA of other species. The screening of milk samples for MRSA showed 47 Positive out of 77 Samples by PCR and 43 positive out of 77 Samples by LAMP. Conclusion: Application of LAMP assay enabled rapid and easy detection of MRSA in milk samples suspected for bovine mastitis. Short Research Article Sathish et al.; BMRJ, 17(1): 1-5, 2016; Article no.BMRJ.28037 2\",\"PeriodicalId\":9269,\"journal\":{\"name\":\"British microbiology research journal\",\"volume\":\"18 1\",\"pages\":\"1-5\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-01-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"British microbiology research journal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.9734/BMRJ/2016/28037\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"British microbiology research journal","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.9734/BMRJ/2016/28037","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1
摘要
目的:建立一种简便、快速的乳腺炎乳样MRSA检测方法。方法:利用现有实验室菌株进行LAMP法的实验室敏感性和特异性分析。牛奶样本采集自印度泰米尔纳德邦的蒂鲁瓦卢尔和坎奇普兰地区。直接从牛奶样品中分离DNA,采用PCR和LAMP方法对mecA基因进行筛选。结果:LAMP法在64°C的等温条件下,在60 min内成功扩增出mecA基因。LAMP法能够检测到10 pg的DNA,并且没有从其他物种的细菌DNA中扩增出mecA基因。77份牛奶样品经PCR检测为阳性47份,经LAMP检测为阳性43份。结论:LAMP法可快速、简便地检测奶牛乳腺炎乳样中的MRSA。研究简述:Sathish et al.;中国生物医学工程学报,17(1):1-5,2016;文章no.BMRJ。28037 2
Rapid Detection of MRSA by Loop Mediated Isothermal Amplification in Bovine Milk Samples
Aim: To develop a simple and rapid detection method for diagnosis of MRSA in bovine milk samples suspected for mastitis. Methodology: The laboratory sensitivity and specificity of LAMP assay was carried out using available laboratory strains. Milk samples were collected from Thiruvallur and Kanchipuram districts of Tamilnadu, India. DNA was directly isolated from the milk samples and mecA gene was screened by both PCR and LAMP methods. Results: The LAMP assay successfully amplified the mecA gene under isothermal conditions at 64°C within 60 min. LAMP assay was able to detect 10 pg of DNA and did not amplify mecA gene from bacterial DNA of other species. The screening of milk samples for MRSA showed 47 Positive out of 77 Samples by PCR and 43 positive out of 77 Samples by LAMP. Conclusion: Application of LAMP assay enabled rapid and easy detection of MRSA in milk samples suspected for bovine mastitis. Short Research Article Sathish et al.; BMRJ, 17(1): 1-5, 2016; Article no.BMRJ.28037 2