title: Detection of Streptococcus pyogenes using rapid visual molecular assay.
abstract: Streptococcus pyogenes is an increasingly important pathogen in many parts of the world. Rapid and accurate detection of S. pyogenes aids in the control of the infection. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of S. pyogenes. The assay incorporates two methods: a chromogenic analysis using a calcein/Mn(2+) complex and real-time turbidity monitoring to assess the reaction. Both methods detected the target DNA within 60 min under 64°C isothermal conditions. The assay used specifically designed primers to target spy1258, and correctly identified 111 strains of S. pyogenes and 32 non-S. pyogenes strains, including other species of the genus Streptococcus. Tests using reference strains showed that the LAMP assay was highly specific. The sensitivity of the assay, with a detection limit of 1.49 pg DNA, was 10-fold greater than that of PCR. The LAMP assay established in this study is simple, fast and sensitive, and does not rely upon any special equipment; thus, it could be employed in clinical diagnosis.
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Xiangna
authors: X
authors: Zhao
authors: College of Food Science and Engineering, Northwest A & F University, Yangling 712100, China.
authors: Xiaoming
authors: X
authors: He
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Huan
authors: H
authors: Li
authors: Emergency Center, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors: Jiangtao
authors: J
authors: Zhao
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Simo
authors: S
authors: Huang
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Wei
authors: W
authors: Liu
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Xiao
authors: X
authors: Wei
authors: Clinical Laboratory, Navy general hospital, Beijing 100048, China.
authors: Yiwei
authors: Y
authors: Ding
authors: Department of Pediatrics, Navy general hospital, Beijing 100048, China.
authors: Zhaoyan
authors: Z
authors: Wang
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Dayang
authors: D
authors: Zou
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Xuesong
authors: X
authors: Wang
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Derong
authors: D
authors: Dong
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Zhan
authors: Z
authors: Yang
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Xiabei
authors: X
authors: Yan
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Liuyu
authors: L
authors: Huang
authors: College of Food Science and Engineering, Northwest A & F University, Yangling 712100, China.
authors: Shuangkui
authors: S
authors: Du
authors: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China yuanjing6216@163.com.
authors: Jing
authors: J
authors: Yuan
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D005452:Fluoresceins; D006801:Humans; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000175:diagnosis; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D005452:Fluoresceins; C007740:fluorexon
keywords: Streptococcus pyogenes; loop-mediated isothermal amplification; spy1258