Comparison of Sybr Green I, ELISA and conventional agarose gel-based PCR in the detection of infectious bursal disease virus

Abstract

The current available molecular method to detect infectious bursal disease virus (IBDV) is by reverse transcriptase-polymerase chain reaction (RT-PCR). However, the conventional PCR is time consuming, prone to error and less sensitive. In this study, the performances of Sybr Green I real-time PCR, enzyme-linked immunosorbent assay (ELISA) and conventional agarose detection methods in detecting specific IBDV PCR products were compared. We found the real-time PCR was at least 10 times more sensitive than ELISA detection method with a detection limit of 0.25pg. The latter was also at least 10 times more sensitive than agarose gel electrophoresis detection method. The developed assay detects both very virulent and vaccine strains of IBDV but not other RNA viruses such as Newcastle disease virus and infectious bronchitis virus. Hence, Sybr Green I-based real-time PCR is a highly sensitive assay for the detection of IBDV.

Comparison of SYBR green I real-time RT-PCR with conventional agarose gel-based RT-PCR for the diagnosis of infectious bronchitis virus infection in chickens in Morocco

“…SYBR green I-based RT-PCR assays have proven to be among the most effective tools in the rapid and differential detection of a variety of viral diseases such as avian influenza, Newcastle disease, and IB. These inexpensive and easily performed assays are important to rapidly identify the causative agent of any upper respiratory disease or changes in egg shell quality and egg production in chickens [ 17 , 18 ].…”

Application of PCR-ELISA in Molecular Diagnosis

“…Several studies compared the use of qPCR, PCR-ELISA, and conventional PCR for the detection of poultry virus, infectious bursal disease virus, and the summary of the results is listed in Table 1 [2, 10–14]. …”

“…Not limiting the use of the method in the medical and food industries, the study of PCR-ELISA extends even to the veterinary industry. Amongst the studies are Leishmania parasite detection [37, 38] and the detection of various avian viruses in chickens [10, 39, 40]. Other detection studies that help detect the presence of plant pathogens [41, 42] include tomato spotted wilt [43], potato spindle tuber [12], prevalence of each phylogenetic group among the infected grapevine varieties [44], and plant viruses in woody plants [45].…”