title: Rapid visual detection of highly pathogenic Streptococcus suis serotype 2 isolates by use of loop-mediated isothermal amplification.
abstract: Streptococcus suis serotype 2 (S. suis 2) is an important zoonotic pathogen that causes considerable economic losses to the pig industry and significantly threatens public health worldwide. The highly pathogenic S. suis 2, which contains the 89K pathogenicity island (PAI), has caused large-scale outbreaks of infections in humans, resulting in high mortality rates. In this study, we established two loop-mediated isothermal amplification (LAMP)-based assays that can rapidly detect S. suis 2 and the 89K PAI and can be performed simultaneously under the same conditions. Further, based on the findings of these two LAMP assays and using the same set of serially diluted DNA samples, we compared the sensitivities of different LAMP product detection methods, including SYBR green detection, gel electrophoresis, turbidimetry, calcein assays, and hydroxynaphthol blue detection. The results suggest that target genes can be amplified and detected within 48 min under 63°C isothermal conditions. The sensitivity of tests for S. suis 2 detection varies between detection methods and reaction systems, indicating that for each LAMP reaction system, multiple detection methods should be performed to select the optimal one. The sensitivities of the optimized methods (7.16 copies/reaction) in the present study were identical to those of the real-time PCR assay, and the test results for reference strains and clinical samples showed that these LAMP systems have high specificities. Thus, since the LAMP systems established in this study are simple, fast, and sensitive, they may have good clinical potential for detecting the highly pathogenic S. suis 2.
authors.affiliation: Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University, Shanghai, China.
authors.forename: Jinhai
authors.initials: J
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Hao
authors.initials: H
authors.lastname: Ren
authors.forename: Shiying
authors.initials: S
authors.lastname: Zhu
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.forename: Fengyu
authors.initials: F
authors.lastname: Zhang
authors.forename: Heng
authors.initials: H
authors.lastname: Lv
authors.forename: Dan
authors.initials: D
authors.lastname: Hu
authors.forename: Lina
authors.initials: L
authors.lastname: Hao
authors.forename: Meiling
authors.initials: M
authors.lastname: Geng
authors.forename: Xiufang
authors.initials: X
authors.lastname: Gong
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Pan
authors.forename: Changjun
authors.initials: C
authors.lastname: Wang
authors.forename: Zhongtian
authors.initials: Z
authors.lastname: Qi
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000175:diagnosis* / Q000382:microbiology*; D017011:Streptococcus suis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*