Comparison of conventional versus real-time PCR detection of Brugia malayi DNA from dried blood spots from school children in a low endemic area.
Microscopic detection of active phase of lymphatic filariasis is indicated by the presence of microfilaria in whole blood. This method is not sensitive and requires relatively large amount of blood sample. PCR allows very sensitive detection of the parasite DNA using a smaller amount of blood; and the use of dried blood spots facilitates sample transportation. Nevertheless, limited studies have been reported on PCR using dried blood spot for detection of Brugia malayi. In this study, we investigated the effects of concentrating whole blood genomic DNA sample and the amplification methods [conventional PCR (C-PCR) and real-time PCR] on the detection of B. malayi DNA from dried blood spots from a very low endemic area in Malaysia. Both C-PCR and real-time PCR detected 2 out of 18 (11%) samples as positive from non-concentrated genomic DNA preparations. After the DNA samples were pooled and concentrated, both C-PCR and realtime PCR detected B. malayi DNA amplifications in 7 out of 18 (39%) samples. However one sample which showed faint band in C-PCR was detected as highly positive in real-time PCR. In conclusion, both C-PCR and real-time PCR using dried blood spots from a low endemic area demonstrated equal sensitivity for detection of B. malayi DNA.
Rahmah
N
N
Institute for Research in Molecular Medicine, Universiti Sains Malaysia (USM), 11800, Pulau Pinang, Malaysia. rahmah8485@gmail.com
Nurulhasanah
O
O
Norhayati
S
S
Zulkarnain
I
I
Norizan
M
M
D000818:Animals; D017178:Brugia malayi / Q000302:isolation & purification*; D002648:Child; D018509:DNA, Helminth / Q000235:genetics*; D019353:Endemic Diseases; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D018509:DNA, Helminth