BACKGROUND: Definitive diagnosis of infestation with Angiostrongylus cantonensis is difficult because the parasitic nematode is undetectable in the cerebrospinal fluid (CSF) of one-half of afflicted patients and the diagnostic sensitivity of ELISA for circulating worm antigens in patient sera is low. We studied immuno-PCR as a diagnostic tool. METHODS: We studied 30 controls and 60 afflicted patients (30 confirmed by parasitologic analysis of CSF). We used a monoclonal antibody to capture circulating A. cantonensis antigens in serum samples. A DNA label generated by PCR amplification with biotinylated primer was bound by use of streptavidin to a biotinylated third antibody. Circulating antigens sandwiched by monoclonal antibody were detected by PCR amplification of the DNA label. RESULTS: The detection limit of the ELISA was 100-1000 times higher than that of the immuno-PCR. The concentrations of circulating antigens in patients were markedly higher than those in controls (Wilcoxon rank-sum test, P <0.001). At a cutoff of 0.1 ng/L, sensitivity and specificity for immunodiagnosis of patients with angiostrongyliasis by immuno-PCR were 98% (95% confidence interval, 91-99%) and 100% (93-100%), respectively. The test was positive in all parasitologically confirmed cases. CONCLUSIONS: Immuno-PCR is a promising technique for diagnosis of A. cantonensis infestation.
BACKGROUND: Definitive diagnosis of infestation with Angiostrongylus cantonensis is difficult because the parasitic nematode is undetectable in the cerebrospinal fluid (CSF) of one-half of afflicted patients and the diagnostic sensitivity of ELISA for circulating worm antigens in patient sera is low. We studied immuno-PCR as a diagnostic tool. METHODS: We studied 30 controls and 60 afflicted patients (30 confirmed by parasitologic analysis of CSF). We used a monoclonal antibody to capture circulating A. cantonensis antigens in serum samples. A DNA label generated by PCR amplification with biotinylated primer was bound by use of streptavidin to a biotinylated third antibody. Circulating antigens sandwiched by monoclonal antibody were detected by PCR amplification of the DNA label. RESULTS: The detection limit of the ELISA was 100-1000 times higher than that of the immuno-PCR. The concentrations of circulating antigens in patients were markedly higher than those in controls (Wilcoxon rank-sum test, P <0.001). At a cutoff of 0.1 ng/L, sensitivity and specificity for immunodiagnosis of patients with angiostrongyliasis by immuno-PCR were 98% (95% confidence interval, 91-99%) and 100% (93-100%), respectively. The test was positive in all parasitologically confirmed cases. CONCLUSIONS: Immuno-PCR is a promising technique for diagnosis of A. cantonensis infestation.
Authors: Yvonne Qvarnstrom; James J Sullivan; Henry S Bishop; Robert Hollingsworth; Alexandre J da Silva Journal: Appl Environ Microbiol Date: 2006-12-28 Impact factor: 4.792
Authors: Ming Jun Deng; Xi Zhi Xiao; Yan Ming Zhang; Xin Hai Wu; Lai Hua Zhu; Xue Qian Xin; Dong Lai Wu Journal: Mol Biol Rep Date: 2010-09-22 Impact factor: 2.316
Authors: Susan I Jarvi; William C Pitt; Margaret E Farias; Laura Shiels; Michael G Severino; Kathleen M Howe; Steven H Jacquier; Aaron B Shiels; Karis K Amano; Blaine C Luiz; Daisy E Maher; Maureen L Allison; Zachariah C Holtquist; Neil T Scheibelhut Journal: PLoS One Date: 2015-04-24 Impact factor: 3.240