Development & evaluation of biotinylated DNA probe for clinical diagnosis of chikungunya infection in patients' acute phase serum & CSF samples
Autor(es): Kumar Jyoti S, Parida Manmohan, Lakshmana Rao P V
Resumo: The resurgence of chikungunya virus (CHIKV) in the Indian Ocean Isl-s - India has drawn worldwide attention due to its explosive nature, high morbidity - complex clinico-pathological manifestations. The early confirmatory diagnosis of CHIKV is essential for management as well as control of unprecedented epidemics. The present study describes the development - evaluation of a highly sensitive - specific E1 structural gene specific biotinylated DNA probe for detection of chikungunya virus in clinical samples using a dot blot format. The complementary DNA (cDNA) of CHIKV was spotted on to nylon membrane. The membrane was subjected to prehybridization - hybridization - developed using a colour development solution containing DAB chromogen. The CHIKV E1 specific DNA probe was highly sensitive detecting picogram levels of target nucleic acid. The comparative evaluation with SYBR Green I based real-time RT-PCR revealed 99 per cent accordance with a sensitivity - specificity of 99 - 98 per cent, respectively. The specificity of this assay was further confirmed through cross-reaction studies with confirmed dengue - Japanese encephalitis (JE) patient serum samples along with infected culture supernatant of Ross River - Saint Louis encephalitis - plasmid DNA of O'Nyong Nyong, Semlinki forest - Sindbis viruses. The DNA probe reported in this study may be useful for specific, sensitive - confirmatory clinical diagnosis of chikungunya infection in acute phase human patient serum - CSF samples. This assay can also be used in the laboratory for quantification of viral antigen in cell culture supernatant for research purpose.
Palavras-Chave: Biotinylated DNA probe; Chikungunya; Clinical diagnosis; E1 structural gene
Imprenta: The Indian Journal of Medical Research, v. 138, p. 117-124, 2013
Descritores: Chikungunya virus - Cell ; Chikungunya virus - DNA ; Chikungunya virus - Viral infections ; Chikungunya virus - PCR detection ; Chikungunya virus - Real Time PCR ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Epidemic ; Chikungunya virus - Public health
Data de publicação: 2013
