Journal for Veterinary Medicine, Biotechnology and Biosafety
Volume
5, Issue 3, September 2019, Pages 19–21
ISSN 2411-3174 (print version) ISSN 2411-0388
(online version)
DEVELOPMENT OF A MULTIPLEX
PCR TEST-SYSTEM FOR DETECTION OF BHV-1, BVDV, CHLAMYDIA SPP. AND MYCOPLASMA SPP.
Isakov M. M.
National Scientific
Center ‘Institute of Experimental and Clinical Veterinary
Medicine’, Kharkiv, Ukraine, e-mail: magomedisakovm@gmail.com
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PDF (print version)
Citation for print version: Isakov, M. M. (2019) ‘Development
of a multiplex PCR test-system for detection of BHV-1, BVDV, Chlamydia spp. and Mycoplasma spp.’, Journal for Veterinary Medicine, Biotechnology and
Biosafety, 5(3), pp. 19–21.
Download
PDF (online version)
Citation for online version: Isakov, M. M. (2019) ‘Development
of a multiplex PCR test-system for detection of BHV-1, BVDV, Chlamydia spp. and Mycoplasma spp.’, Journal for Veterinary Medicine, Biotechnology and
Biosafety. [Online] 5(3), pp. 19–21. DOI: 10.36016/JVMBBS-2019-5-3-4.
Summary. This study describes development of a multiplex PCR
assay for detection of BHV-1, BVDV, Chlamydia spp.
and Mycoplasma spp. infections
in bovines. The assay was developed using genomic DNA and RNA and four sets of
PCR primers targeting 16S rRNA genes of Chlamydia spp.,
Mycoplasma spp., 5’-UTR of
Bovine viral diarrhea virus, gE of Bovine herpesvirus-1, respectively. A total
of 100 tissue samples were collected from cattle suspected to be infected
with the viral and bacterial pathogens (BVDV, BHV-1, Chlamydia spp. and Mycoplasma spp.)
from different regions of Ukraine. A part of sample was stored at
–50°C for isolation of genomic DNA and RNA. The multiplex PCR assay
was optimized in the study. The specific primers designed and used in the study
were found sensitive and specific in amplifying target genes viz. 16S rRNA, gE,
5’-UTR of Chlamydia spp., Mycoplasma spp., BHV-1 and BVDV,
respectively. The PCR primers used in the optimization of multiplex PCR assay
for detection of Bovine viral diarrhea virus, Bovine herpesvirus-1, Chlamydia spp., Mycolasma spp. could amplify 221 bp, 111 bp,
386 bp, 279 bp products, respectively. Non specific amplification was
not observed
Keywords: Bovine herpesvirus-1, Bovine viral diarrhoea
virus, Chlamydia, Mycoplasma, multiplex PCR, bovines
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