Development and evaluation of antigen capture ELISA for early clinical diagnosis of chikungunya
Autor(es): Shukla Jyoti, Khan Mohsin, Tiwari Mugdha, Sannarangaiah Santhosh, Sharma Shashi, Rao Putcha Venkata Lakshmana, Parida Manmohan
Resumo: The resurgence of chikungunya (CHIK) in the form of unprecedented explosive epidemic after a gap of 3 decades in India - Indian Ocean isl-s is a point of major public health concern. The laboratory diagnosis is essentially based on virus isolation, IgM ELISA, - reverse transcriptase polymerase chain reaction (RT-PCR). Although PCR-based methods are used for early - accurate diagnosis, the high cost of the assay - requirement of thermal cycler limit its application only to referral laboratories. The antibody-based IgM ELISA is found to be cost-effective, but it takes 5 to 6 days for the patient to develop antibody -, thus, has less implication for early clinical diagnosis - patient management. Therefore, a simple rapid, sensitive, - specific antigen detection system is reported for early - reliable clinical diagnosis as well as effective surveillance of CHIK. A double antibody s-wich system was designed for antigen capture ELISA, employing rabbit - mouse anti-CHIK IgG antibodies as capture - detector antibodies, respectively. An optimal assay condition with 0 background was established having no reactivity with healthy human serum - Cerebro spinal fluid (CSF) samples. The comparative evaluation with SYBR Green I-based real-time RT-PCR revealed an accordance of 96% with a sensitivity - specificity of 95% - 97%, respectively. The specificity of this assay was confirmed through cross-reactivity studies with confirmed dengue - Japanese encephalitis (JE) patient serum - CSF samples. The antigen capture ELISA reported in this study was able to detect the presence of viral antigen as early as the second day of fever -, thus, can be very useful for early clinical diagnosis of CHIK with acute phase patient serum - CSF samples. This can also be used for rapid screening of large numbers of clinical samples in endemic areas during epidemics.
Imprenta: Diagnostic Microbiology and Infectious Disease, v. 65, n. 2, p. 142-149, 2009
Identificador do objeto digital: 10.1016/j.diagmicrobio.2009.06.017
Descritores: Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Antibodies ; 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 - Immunology ; Chikungunya virus - Public health
Data de publicação: 2009
