Evaluation of PCR methods for detection of Brucella strains from culture and tissues
| Autor: | Alper Çiftci, B. Sareyyüpoğlu, Mehmet Akan, K. S. Diker, Serap Savaşan, Tuba Iça |
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| Přispěvatelé: | Ondokuz Mayıs Üniversitesi |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
DNA Bacterial 040301 veterinary sciences 030106 microbiology Sheep Diseases Semen Brucella Polymerase Chain Reaction Sensitivity and Specificity Brucellosis Microbiology 0403 veterinary science 03 medical and health sciences Food Animals Species Specificity Pregnancy RNA Ribosomal 16S Zoonoses medicine Animals Sheep Domestic DNA Primers Bacteriological Techniques Sheep biology Inoculation Zoonosis 04 agricultural and veterinary sciences Genus Brucella Abortion Veterinary biology.organism_classification medicine.disease 16S ribosomal RNA Isolation (microbiology) Virology Detection PCR Milk Aborted Fetus Animal Science and Zoology Cattle Female |
| Zdroj: | Tropical animal health and production. 49(4) |
| ISSN: | 1573-7438 |
| Popis: | Akan, Mehmet/0000-0002-7342-1450 WOS: 000399021800012 PubMed: 28255651 The genus Brucella causes significant economic losses due to infertility, abortion, stillbirth or weak calves, and neonatal mortality in livestock. Brucellosis is still a zoonosis of public health importance worldwide. The study was aimed to optimize and evaluate PCR assays used for the diagnosis of Brucella infections. For this aim, several primers and PCR protocols were performed and compared with Brucella cultures and biological material inoculated with Brucella. In PCR assays, genus- or species-specific oligonucleotide primers derived from 16S rRNA sequences (F4/R2, Ba148/928, IS711, BruP6-P7) and OMPs (JPF/JPR, 31ter/sd) of Brucella were used. All primers except for BruP6-P7 detected the DNA from reference Brucella strains and field isolates. In spiked blood, milk, and semen samples, F4-R2 primer-oriented PCR assays detected minimal numbers of Brucella. In spiked serum and fetal stomach content, Ba148/928 primer-oriented PCR assays detected minimal numbers of Brucella. Field samples collected from sheep and cattle were examined by bacteriological methods and optimized PCR assays. Overall, sensitivity of PCR assays was found superior to conventional bacteriological isolation. Brucella DNA was detected in 35.1, 1.1, 24.8, 5.0, and 8.0% of aborted fetus, blood, milk, semen, and serum samples by PCR assays, respectively. In conclusion, PCR assay in optimized conditions was found to be valuable in sensitive and specific detection of Brucella infections of animals. Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [VHAG-1902] This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) (Project No: VHAG-1902). |
| Databáze: | OpenAIRE |
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