scholarly journals COMPARISON OF AGAR GEL IMMUNODIFFUSION TEST, IMMUNOHISTOCHEMSITRY AND REVERSE TRANSCRIPTION - POLYMERASE CHAIN REACTION FOR DETECTION OF INFECTIOUS BURSAL DISEASE VIRUS

2013 ◽  
Vol 9 (2) ◽  
pp. 121-125 ◽  
Author(s):  
MT Islam ◽  
MN Islam ◽  
MZI Khan ◽  
MA Islam
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Infectious Bursal Disease Virus ◽  
Infectious Bursal Disease ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Agar Gel ◽  
Bursal Disease ◽  
Polymerase Chain

The objective of this study was to compare agar gel immunodiffusion test (AGID) and immunohistochemistry (IHC) with reverse transcriptase-polymerase chain reaction (RT-PCR) in terms of sensitivity and specificity for the detection of infectious bursal disease virus (IBDV). Thirty-five bursal samples collected from filed outbreak of IBD were evaluated by all 3 diagnostic tests. Sensitivity and specificity of both AGIDT and IHC with RT-PCR was 94.12% and 100%, respectively. Both AGIDT and IHC showed a 94.29% association with RT-PCR with a k value of 0.482, indicating a moderate degree of agreement. The Cochran’s Q value was 4.00, which is lower than the critical value, indicating that the methods did not differ significantly (p>0.05) from each other in detection of samples as positive or negative. However, RT-PCR had distinct edge over these two tests employed in the study.DOI: http://dx.doi.org/10.3329/bjvm.v9i2.13452

scholarly journals Development of multiplex reverse transcription-polymerase chain reaction for differentiation of strains of infectious bursal disease virus and primary screening of the virus in Thailand

2021 ◽  
pp. 3105-3110
Author(s):  
Nataya Charoenvisal
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Reverse Transcription ◽  
Infectious Bursal Disease Virus ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Bursal Disease ◽  
Polymerase Chain

Background and Aim: A new set of primers (400 base pairs partial of VP2) was designed and used for the infectious bursal disease virus (IBDV) screening test. Using this new primer set, the enzymes MboI and BstNI were unable to differentiate the field and vaccine strains. As a result, a new simple, cheap, and appropriate tool for strain differentiation is required. The objective of this study was to develop the appropriate restriction fragment length polymorphism (RFLP) and multiplex reverse transcription-polymerase chain reaction (RT-PCR) for the differentiation of classic IBDV (cIBDV) strains and very virulent IBDV (vvIBDV) strains in Thailand. Materials and Methods: Ninety seven bursa of Fabricius from 16 farms were collected from farms in the eastern and central regions of Thailand. RT-PCR screening showed that 82 samples were positive for IBDV and 15 samples were negative. Then, selected samples were sequenced from each farm with a positive test. Results: The sequencing results showed that samples from six of the farms were vvIBDV and samples from the other six farms were cIBDV. Although the whole genome sequencing was incomplete, both the sequencing results of segment A and segment B showed high similarity between cIBDV and vvIBDV. Restriction enzyme cutting site and primers for multiplex RT-PCR were hard to design. An RT-PCR-RFLP method was developed, but it failed to differentiate IBDV strains. However, the multiplex RT-PCR was able to differentiate cIBDV from vvIBDV. Four primers were used in the multiplex RT-PCR. Conclusion: These four primers were used together in one reaction at an annealing temperature of 45°C. Therefore, multiplex RT-PCR is a less complicated, cheaper, and less time-consuming method for the differentiation of cIBDV and vvIBDV strains.

Sign in / Sign up

Export Citation Format

Share Document