Publication: Polymerase Chain Reaction Of Dried Blood Spots To Detect Parasite DNA In Individuals With Lymphatic Filariasis
Date
1999
Authors
Sabudin, Mazula
Journal Title
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Abstract
Lymphatic filariasis caused by Brugia malayi has traditionally been detected
in the blood of infected individuals by microscopy. Screening for blood-stage
microfilaria (mf) by microscopy is labour intensive with user fatigue and poor
specimen handling responsible for false negative results. Recently a method
to detect the DNA from circulating microfilaria using the polymerase chain
reaction (PCR) has been described (Lizotte et a/., 1994 ). However, the
specimen collection method described \Yas unsuitable for routine screening in
· field situations. The aim of the study reported here was to adapt the PCR
method to a simple blood spot sampling and DNA extraction method suitable
for remote areas without compromising the sensitivity of PCR. Blood spots
were collected from individuals in Kelantan and Terengganu to optimise the
technique. A one tube DNA extraction method was developed and coupled to
a nested PCR assay that was field tested on an endemic community in
Sabah. There was 100°/o sensitivity when comparing PCR to microscopy but
only 70% sensitivity when comparing microscopy to PCR. The increased
sensitiyity of PCR coupled with simple sample collection and DNA extraction
provides a valuable ~ltern~tive to microscopy for detecting B. ma/ayi positive
individuals in endem1c reg1ons of the world. ·