| Title of Article |
Rapid detection of Wuchereria bancrofti and Brugia malayi in mosquito vectors (Diptera: Culicidae) using a real-time fluorescence resonance energy transfer multiplex PCR and melting curve analysis. |
| Date of Acceptance |
3 October 2008 |
| Journal |
| Title of Journal |
Journal of medical entomology |
| Standard |
|
| Institute of Journal |
B.P. Bishop Museum, Entomology Dept. |
| ISBN/ISSN |
|
| Volume |
|
| Issue |
46 |
| Month |
มกราคม |
| Year of Publication |
2010 |
| Page |
158-164 |
| Abstract |
ABSTRACT We developed a single-step real-time ßuorescence resonance energy transfer (FRET)
multiplex polymerase chain reaction (PCR) merged with melting curve analysis for the detection of
Wuchereria bancrofti and Brugia malayiDNAin blood-fed mosquitoes. Real-timeFRETmultiplexPCR
is based on ßuorescence melting curve analysis of a hybrid of amplicons generated from two families
of repeatedDNAelements: the 188 bp SspI repeated sequence, speciÞc to W. bancrofti, and the 153-bp
HhaI repeated sequence, speciÞc to the genus Brugia and two pairs of speciÞc ßuorophore-labeled
probes. Both W. bancrofti and B. malayi can be differentially detected in infected vectors by this
process through their different ßuorescence channel and melting temperatures. The assay could
distinguish both human Þlarial DNAs in infected vectors from the DNAs of Dirofilaria immitis– and
Plasmodium falciparumÐinfected human red blood cells and noninfected mosquitoes and human
leukocytes. The technique showed 100% sensitivity and speciÞcity and offers a rapid and reliable
procedure for differentially identifying lymphatic Þlariasis. The introduced real-time FRET multiplex
PCR can reduce labor time and reagent costs and is not prone to carry over contamination. The test
can be used to screen mosquito vectors in endemic areas and therefore should be a useful diagnostic
tool for the evaluation of infection rate of the mosquito populations and for xenomonitoring in the
community after eradication programs such as the Global Program to Eliminate Lymphatic Filariasis. |
| Keyword |
Wuchereria bancrofti , Brugia malayi,Detection, Mosquito, Real-time PCR |
| Author |
|
| Reviewing Status |
มีผู้ประเมินอิสระ |
| Status |
ตีพิมพ์แล้ว |
| Level of Publication |
นานาชาติ |
| citation |
false |
| Part of thesis |
true |
| Attach file |
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| Citation |
0
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Publication
Journal Publication
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