Article ; Online: Comparison of bacteriological culture method and multiplex real-time PCR for detection of mastitis.
Research in veterinary science
2024 Volume 172, Page(s) 105237
Abstract: This study includes the evaluation of multiplex real-time PCR (rPCR) kit, which was developed to provide rapid diagnosis of mastitis infections, by working with milk samples of 2 different sources of mastitis and comparing the results with the classical ... ...
| Abstract | This study includes the evaluation of multiplex real-time PCR (rPCR) kit, which was developed to provide rapid diagnosis of mastitis infections, by working with milk samples of 2 different sources of mastitis and comparing the results with the classical bacteriological culture method (BC). A total of 273 bacteria were isolated in 226 samples (47.88%) out of 472 samples by BC. These were 139 (50.91%) Staphylococcus spp., 61 (22.34%) Streptococcus spp., 15 (5.49%) E. coli, 8 (2.93%) Enterococcus spp., 50 (18.31%) other bacteria. When we look at the multiplex rPCR results; 1052 positive were obtained for the gene regions of 14 different bacteria, 1 yeast, and 1 β-lactamase gene examined in 472 samples. While no searched gene region was found by rPCR in 78 (16.5%) of the 472 samples studied, at least 1 gene was detected in 394 (83.5%) samples. These 1052 positive samples by rPCR were; 263 (28.43%) Staphylococcus spp., 51 (5.51%) S. aureus, 57 (6.16%) Enterococcus spp., 49 (5.29%) C. bovis, 16 (1.73%) S. dysgalactiae, 84 (9.08%) S. agalactiae, 71 (7.67%) S. uberis, 73 (7.89%) E. coli, 14 (1.51%) Prototheca spp., 39 (4.21%) T. pyogenes/P. indolicus, 5 (0.54%) S. marcescens, 15 (1.62%) K. oxytoca/pneumonia, 117 (12.64%) Mycoplasma spp., 31 (3.35%) M. bovis, 40 (4.32%) yeast, and 127 samples (26.90%) were β-lactamase positive. When the antibiotic resistance of the isolates was evaluated, 78 (31.96%) tetracycline, 72 (29.5%) penicillin, and 60 (24.59%) clindamycin resistance were observed predominantly in Gram-positive isolates, while 6 (23.07%) tigecycline, 6 (23.07%) netilmicin, 6 (23.07%) pipercillin resistance was found in gram-negative isolates. While a bacteria and/or yeast gene was found by rPCR in 187 of 246 (76.01%) samples with no bacterial growth, a bacterium was isolated with BC in only 20 (8.84%) samples whose gene region was not found by rPCR. As a result, the multiplex rPCR system used in the diagnosis of mastitis has been found to be quite reliable as it can detect a large number of bacteria in a very short time compared to classical methods. Therefore, we advise the use of rPCR and/or culture for confirmation of clinical signs in mastitis and at routine mastitis surveillance. |
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| MeSH term(s) | Mastitis, Bovine/microbiology ; Mastitis, Bovine/diagnosis ; Female ; Animals ; Real-Time Polymerase Chain Reaction/veterinary ; Real-Time Polymerase Chain Reaction/methods ; Cattle ; Multiplex Polymerase Chain Reaction/veterinary ; Multiplex Polymerase Chain Reaction/methods ; Bacteriological Techniques/methods ; Bacteriological Techniques/veterinary ; Milk/microbiology ; Bacteria/isolation & purification ; Bacteria/genetics ; Bacteria/classification |
| Language | English |
| Publishing date | 2024-03-22 |
| Publishing country | England |
| Document type | Journal Article ; Comparative Study |
| ZDB-ID | 840961-4 |
| ISSN | 1532-2661 ; 0034-5288 |
| ISSN (online) | 1532-2661 |
| ISSN | 0034-5288 |
| DOI | 10.1016/j.rvsc.2024.105237 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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