Publication:
High resolution melting real-time PCR for rapid discrimination between Brugia malayi and Brugia pahangi.

2
0

Issued Date

2009

Resource Type

Article

File Type

application/pdf

ISSN

1252208

Other identifier(s)

2-s2.0-77957315224

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of the Medical Association of Thailand = Chotmaihet thangphaet. Vol 92 Suppl 3, (2009), p.S24-28

Suggested Citation

Areekit S., Kanjanavas P., Pakpitchareon A., Khawsak P., Khuchareontaworn S., Sriyaphai T., Chansiri K. High resolution melting real-time PCR for rapid discrimination between Brugia malayi and Brugia pahangi.. Journal of the Medical Association of Thailand = Chotmaihet thangphaet. Vol 92 Suppl 3, (2009), p.S24-28. Retrieved from: https://hdl.handle.net/20.500.14740/7693

Title

High resolution melting real-time PCR for rapid discrimination between Brugia malayi and Brugia pahangi.

Author(s)

Areekit S.
 Kanjanavas P.
 Pakpitchareon A.
 Khawsak P.
 Khuchareontaworn S.
 Sriyaphai T.
 Chansiri K.

Abstract

OBJECTIVE: To identify two closely related Brugia malayi and B. pahangi in cat reservoirs by using high resolution melting real-time PCR (HRM real-time PCR). MATERIAL AND METHOD: HRM analysis on the Corbett Rotor-Gene 6000 instrument was used to test 5 Brugia specimens by using five sets of specific primers for HhaI repetitive region (HR), small heat shock protein (SHP), small subunit ribosomal DNA (18S rDNA), internal transcribed spacer region (ITS), and trans-spliced leading Exon I gene (SLX1). RESULTS: HRM analysis of ITS and SLX clearly generated 2 profiles of B. malayi and B. pahangi while those of HR, 18S rDNA, and SHP could classify B. pahangi. CONCLUSION: HRM is a simple and rapid method for identification of two closely related B. malayi and B. pahangi in which it can detect both parasites within 30 min after real-time PCR detection. This assay is probe-free HRM and reduces a risk of PCR carryover. It does not require multiplex methods and DNA sequencing; therefore, HRM provides a new approach for genetic screening and rapid detection of closely related species in a clinical laboratory.

Subject(s)

Heat shock protein
 Animal
 Article
 Brugia malayi
 Brugia pahangi
 Cat
 DNA microarray
 DNA sequence
 Genetics
 Human
 Instrumentation
 Isolation and purification
 Nucleotide repeat
 Reverse transcription polymerase chain reaction
 Time
 Animals
 Brugia malayi
 Brugia pahangi
 Cats
 Heat-Shock Proteins
 Humans
 Oligonucleotide Array Sequence Analysis
 Repetitive Sequences, Nucleic Acid
 Reverse Transcriptase Polymerase Chain Reaction
 Sequence Analysis, DNA
 Time Factors

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https://hdl.handle.net/20.500.14740/7693

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Scopus: Year 1983-2021

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