Comparing the performance of conventional PCR, RTQ-PCR, and droplet digital PCR assays in detection of Shigella.
作者： Jin Yang , Nana Zhang , Jun Lv , Ping Zhu , Xing Pan
DOI: 10.1016/J.MCP.2020.101531
关键词: genomic DNA 、 Polymerase chain reaction 、 Molecular biology 、 Shigella 、 Quantification methods 、 Digital polymerase chain reaction 、 Quantitative Real Time PCR 、 Detection limit 、 Microbiological culture 、 Biology
摘要: Abstract The incidence of foodborne infections caused by Shigella spp. is still very high in every year, which poses a great potential threat to public health. Conventional quantification methods based on culture techniques, biochemical, and serological identification are time-consuming labor-intensive. To develop more rapid efficient detection method spp., we compared the sensitivity specificity three different polymerase chain reaction (PCR) methods, including conventional PCR, quantitative real-time PCR (RTQ-PCR), droplet digital (ddPCR). Our results indicated that ddPCR exhibited higher sensitivity, limit was 10−5 ng/μl for genomic DNA templates, 10−1 cfu/ml bacteria culture. In addition, found time-saving method, required shorter pre-culturing time. Collectively, assay reliable effective
sci-hub.se 参考文章(32)
M. Pavlovic, A. Luze, R. Konrad, A. Berger, A. Sing, U. Busch, I. Huber, Development of a duplex real‐time PCR for differentiation between E. coli and Shigella spp. Journal of Applied Microbiology. ,vol. 110, pp. 1245- 1251 ,(2011) , 10.1111/J.1365-2672.2011.04973.X
P. Van Lint, E. De Witte, H. De Henau, A. De Muynck, L. Verstraeten, B. Van Herendael, S. Weekx, Evaluation of a real-time multiplex PCR for the simultaneous detection of Campylobacter jejuni, Salmonella spp., Shigella spp./EIEC, and Yersinia enterocolitica in fecal samples. European Journal of Clinical Microbiology & Infectious Diseases. ,vol. 34, pp. 535- 542 ,(2015) , 10.1007/S10096-014-2257-X
Il-Hoon Cho, Jin-Woo Jeon, Sung-Ho Paek, Dong-Hyung Kim, Hee-Sung Shin, Un-Hwan Ha, Sung-Kyu Seo, Se-Hwan Paek, Toll-like receptor-based immuno-analysis of pathogenic microorganisms. Analytical Chemistry. ,vol. 84, pp. 9713- 9720 ,(2012) , 10.1021/AC300668Y
María R. Viñas, Ezequiel Tuduri, Alicia Galar, Katherine Yih, Mariana Pichel, John Stelling, Silvina P. Brengi, Anabella Della Gaspera, Claudia van der Ploeg, Susana Bruno, Ariel Rogé, María I. Caffer, Martin Kulldorff, Marcelo Galas, , Laboratory-Based Prospective Surveillance for Community Outbreaks of Shigella spp. in Argentina PLOS Neglected Tropical Diseases. ,vol. 7, ,(2013) , 10.1371/JOURNAL.PNTD.0002521
A.I. Camacho, J. Souza-Rebouças, J.M. Irache, C. Gamazo, Towards a non-living vaccine against Shigella flexneri: from the inactivation procedure to protection studies. Methods. ,vol. 60, pp. 264- 268 ,(2013) , 10.1016/J.YMETH.2012.09.008
Divya T. George, Carolyn A. Behm, David H. Hall, Ulrike Mathesius, Melanie Rug, Ken C. Q. Nguyen, Naresh K. Verma, Shigella flexneri Infection in Caenorhabditis elegans: Cytopathological Examination and Identification of Host Responses PLoS ONE. ,vol. 9, pp. e106085- ,(2014) , 10.1371/JOURNAL.PONE.0106085
Fitzroy A Orrett, Prevalence of Shigella serogroups and their antimicrobial resistance patterns in southern Trinidad. Journal of Health Population and Nutrition. ,vol. 26, pp. 456- 462 ,(2009) , 10.3329/JHPN.V26I4.1889
Christopher M Hindson, John R Chevillet, Hilary A Briggs, Emily N Gallichotte, Ingrid K Ruf, Benjamin J Hindson, Robert L Vessella, Muneesh Tewari, Absolute quantification by droplet digital PCR versus analog real-time PCR Nature Methods. ,vol. 10, pp. 1003- 1005 ,(2013) , 10.1038/NMETH.2633
Wen Luo, Sanying Wang, Xuanxian Peng, Identification of shiga toxin-producing bacteria by a new immuno-capture toxin gene PCR. Fems Microbiology Letters. ,vol. 216, pp. 39- 42 ,(2002) , 10.1111/J.1574-6968.2002.TB11411.X
R. Lindqvist, Detection of Shigella spp. in food with a nested PCR method – sensitivity and performance compared with a conventional culture method Journal of Applied Microbiology. ,vol. 86, pp. 971- 978 ,(1999) , 10.1046/J.1365-2672.1999.00777.X