Detection and differentiation of filarial parasites by universal primers and polymerase chain reaction-restriction fragment length polymorphism analysis

Surang Nuchprayoon, Alisa Junpee, Yong Poovorawan, Alan Leroy Scott

Research output: Contribution to journalArticle

Abstract

Filarial nematode parasites are a serious cause of morbidity in humans and animals. Identification of filarial infection using traditional morphologic criteria can be difficult and lead to misdiagnosis. We report on a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based method to detect and differentiate a broad range of filarial species in a single PCR. The first internal transcribed spacer 1 (ITS1) along with the flanking 18S and 5.8S ribosomal DNA (rDNA) were isolated and cloned from Wuchereria bancrofti, Brugia malayi, and Brugia pahangi. Sequence analysis identified conserved sites in the 18S and 5.8S rDNA sequence that could be used as universal priming sites to generate ITS1-distinctive PCR products that were useful for distinguishing filariae at the genus level. The addition of a digestion of the ITS1 PCR product with the restriction endonuclease Ase 1 generated a fragment profile that allowed differentiation down to the species level for W. bancrofti, B. malayi, B. pahangi, Dirofilaria immitis, and D. repens. The PCR-RFLP of ITS1 rDNA will be useful in diagnosing and differentiating filarial parasites in human, animal reservoir hosts, and mosquito vectors in disease-endemic areas.

Original languageEnglish (US)
Pages (from-to)895-900
Number of pages6
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume73
Issue number5
StatePublished - Nov 2005

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Restriction Fragment Length Polymorphisms
Parasites
Polymerase Chain Reaction
Brugia pahangi
Brugia malayi
Wuchereria bancrofti
Ribosomal DNA
Ribosomal Spacer DNA
Dirofilaria immitis
Endemic Diseases
DNA Restriction Enzymes
Diagnostic Errors
Sequence Analysis
Digestion
Morbidity
Infection

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Detection and differentiation of filarial parasites by universal primers and polymerase chain reaction-restriction fragment length polymorphism analysis. / Nuchprayoon, Surang; Junpee, Alisa; Poovorawan, Yong; Scott, Alan Leroy.

In: American Journal of Tropical Medicine and Hygiene, Vol. 73, No. 5, 11.2005, p. 895-900.

Research output: Contribution to journalArticle

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