Paper chromatography hybridization: a rapid method for detection of Onchocerca volvulus DNA amplified by PCR.
Prior studies have shown that Onchocerca volvulus DNA can be detected in skin snips and in black flies after polymerase chain reaction (PCR) with primers specific for repeated "O-150" DNA sequences. We have adapted a paper chromatography hybridization assay (PCHA) to detect amplified O-150 DNA and compared this method to two established methods, namely agarose gel electrophoresis (AGE) and hybridization enzyme-linked immunosorbent assay (ELISA). The minimum amounts of purified O-150 DNA detected by PCHA, AGE, and ELISA were 5, 10, and 2 ng, respectively. The three methods had similar estimated sensitivities for detecting O. volvulus DNA amplified from skin snips from African subjects with onchocerciasis (88%, 84%, and 91%, respectively). No false positive results were observed with skin snips from uninfected control subjects. The paper chromatography hybridization assay detects PCR products in 30 minutes without electricity or special equipment. This technology brings DNA detection a step closer to widespread use in field settings.
Zhang
S
S
Department of Internal Medicine, Barnes-Jewish Hospital and Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Li
B W
BW
Weil
G J
GJ
D000818:Animals; D002163:Cameroon; D016022:Case-Control Studies; D002854:Chromatography, Paper / Q000592:standards*; D017931:DNA Primers; D018509:DNA, Helminth / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005869:Ghana; D006801:Humans; D009693:Nucleic Acid Hybridization; D017181:Onchocerca volvulus / Q000235:genetics*; D009855:Onchocerciasis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology; D012876:Skin Diseases, Parasitic / Q000175:diagnosis*
D017931:DNA Primers; D018509:DNA, Helminth