Publication:
Detection of pathogenic leptospires by loop-mediated isothermal amplification targeting lipL32 Gene

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Issued Date

2015

Resource Type

Article

File Type

application/pdf

ISSN

1252208

Other identifier(s)

2-s2.0-84957659820

Rights Holder(s)

มหาวิทยาลัยศรีนครินทรวิโรฒ

Bibliographic Citation

Journal of the Medical Association of Thailand. Vol 98, (2015), p.S78-S84

Suggested Citation

Seesom W., Thongket P., Rattanathanawan K., Mekseepralard C., Sukhumsirichar W. Detection of pathogenic leptospires by loop-mediated isothermal amplification targeting lipL32 Gene. Journal of the Medical Association of Thailand. Vol 98, (2015), p.S78-S84. Retrieved from: https://hdl.handle.net/20.500.14740/6058

Title

Detection of pathogenic leptospires by loop-mediated isothermal amplification targeting lipL32 Gene

Author(s)

Seesom W.
 Thongket P.
 Rattanathanawan K.
 Mekseepralard C.
 Sukhumsirichar W.

Abstract

Background: Leptospirosis is a worldwide re-emerging infectious disease caused by pathogenic leptospires including Leptospira interrogans. Objective: In the present study, a loop-mediated isothermal amplification (LAMP) was developed to detect L. interrogans using lipL32 as a gene target. Material and Method: Four specific primers were designed based on the conserved region of lipL32 gene of various serovars of pathogenic leptospires. LAMP reaction was performed at 65°C for 1 hour. The LAMP products were detected by agarose gel electrophoresis and fluorescence dye. Results: The lipL32 LAMP assay showed highly specificity to the reference stains of L. interrogans serovar Autumnalis, Bataviae, Javanica, Pyrogenes, Icterohaemorrhagiae, and Saigon. No product was produced from non-pathogenic leptospire (L. biflexa), human, or Escherichia coli. The lower limit of detection analyzed by agarose gel electrophoresis and fluorescence dye visualization was 0.02 pg/μl which equivalent to 4 genomic equivalents/reaction. Moreover, the clinical strain of leptospires including pathogenic and intermediate group of L. interrogans were detected by lipL32 LAMP. Conclusion: The developed lipL32 LAMP is high specificity and sensitivity that can be applied to detect pathogenic leptospires in clinical samples. © 2015, Medical Association of Thailand. All rights reserved.

Subject(s)

Fluorescent dye
 Genomic DNA
 Primer DNA
 Agar gel electrophoresis
 Article
 Bacterial gene
 Bacterial growth
 Bacterium detection
 Clinical article
 Controlled study
 DNA sequence
 Genetic similarity
 Human
 Leptospira interrogans
 Leptospirosis
 Limit of detection
 LipL32 gene
 Loop mediated isothermal amplification
 Polymerase chain reaction
 Sensitivity and specificity
 Serotype
 Genetics
 Leptospira
 Leptospirosis
 Nucleic acid amplification
 Procedures
 DNA Primers
 Humans
 Leptospira
 Leptospirosis
 Nucleic Acid Amplification Techniques
 Sensitivity and Specificity

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URI

https://hdl.handle.net/20.500.14740/6058

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Scopus: Year 1983-2021

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