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Validation of real-time polymerase chain reaction versus conventional polymerase chain reaction for diagnosis of brucellosis in cattle sera
- Source :
- Veterinary World, Veterinary World, Vol 14, Iss 1, Pp 144-154 (2021)
- Publication Year :
- 2021
- Publisher :
- Veterinary World, 2021.
-
Abstract
- Background and Aim: Different polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real- Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera. Materials and Methods: One hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard. Results: TaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p
- Subjects :
- Veterinary medicine
Brucella
Biology
SF1-1100
Serology
law.invention
03 medical and health sciences
0302 clinical medicine
law
SF600-1100
medicine
TaqMan
030212 general & internal medicine
diagnostic sensitivity
Polymerase chain reaction
0303 health sciences
General Veterinary
Receiver operating characteristic
030306 microbiology
Brucellosis
Gold standard (test)
biology.organism_classification
medicine.disease
Virology
Animal culture
Real-time polymerase chain reaction
diagnostic specificity
TaqMan real-time-polymerase chain reaction
bacteriological results
conventional polymerase chain reaction
Research Article
Subjects
Details
- ISSN :
- 22310916 and 09728988
- Volume :
- 14
- Database :
- OpenAIRE
- Journal :
- January-2021
- Accession number :
- edsair.doi.dedup.....5b6bdf2859e5fd703a455513d36f9af2