title: A Comparison of In-House Real-Time LAMP Assays with a Commercial Assay for the Detection of Pathogenic Bacteria.
abstract: Molecular detection of bacterial pathogens based on LAMP methods is a faster and simpler approach than conventional culture methods. Although different LAMP-based methods for pathogenic bacterial detection are available, a systematic comparison of these different LAMP assays has not been performed. In this paper, we compared 12 in-house real-time LAMP assays with a commercialized kit (Isothermal Master Mix) for the detection of Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121, E. coli O145 and Streptococcus agalactiae. False-positive results were observed in all 12 in-house real-time LAMP assays, while all the negative controls of Isothermal Master Mix remained negative after amplification. The detection limit of Isothermal Master Mix for Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121 and Streptococcus agalactiae was 1 pg, whereas the sensitivity of the commercialized kit for E. coli O145 was 100 pg. In conclusion, the 12 in-house real-time LAMP assays were impractical to use, while the commercialized kit Isothermal Master Mix was useful for the detection of most bacterial pathogens.
authors.affiliation: Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China. wangdg666@126.com.
authors.forename: Deguo
authors.initials: D
authors.lastname: Wang
authors.affiliation: Public Lab Center, Xuchang University, Xuchang 461000, Henan, China. wangyzhxuch@sina.com.
authors.forename: Yongzhen
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China. xfug@163.com.
authors.forename: Fugang
authors.initials: F
authors.lastname: Xiao
authors.affiliation: Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China. gwy2002@126.com.
authors.forename: Weiyun
authors.initials: W
authors.lastname: Guo
authors.affiliation: Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China. zyq336@163.com.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Life Science College, Zhengzhou University, Zhengzhou 450000, Henan, China. pingaw@126.com.
authors.forename: Aiping
authors.initials: A
authors.lastname: Wang
authors.affiliation: Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, PA 19038, USA. wangdg666@126.com.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Liu
mesh_terms: D015373:Bacterial Typing Techniques / Q000295:instrumentation / Q000379:methods*; D017931:DNA Primers / Q000138:chemical synthesis; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D005189:False Positive Reactions; D005516:Food Microbiology; D006801:Humans; D057230:Limit of Detection; D008089:Listeria monocytogenes / Q000235:genetics* / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D013211:Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification; D013292:Streptococcus agalactiae / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic
keywords: pathogenic bacteria; real-time loop-mediated isothermal amplification (LAMP); sensitivity; specificity