Please use this identifier to cite or link to this item: https://ir.swu.ac.th/jspui/handle/123456789/13639
ชื่อเรื่อง: Detection of pathogenic leptospires by loop-mediated isothermal amplification targeting lipL32 Gene
ผู้แต่ง: Seesom W.
Thongket P.
Rattanathanawan K.
Mekseepralard C.
Sukhumsirichar W.
Keywords: 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
วันที่เผยแพร่: 2015
บทคัดย่อ: 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.
URI: https://ir.swu.ac.th/jspui/handle/123456789/13639
https://www.scopus.com/inward/record.uri?eid=2-s2.0-84957659820&partnerID=40&md5=fbbb6329f68d1853a77031c75f9065f8
ISSN: 1252208
Appears in Collections:Scopus 1983-2021

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