A novel polymerase chain reaction assay for the detection of seven Mycoplasma species of cattle origin

The study aimed to develop a pair of polymerase chain reaction primers for detecting ruminant mycoplasma pathogens. We designed a set of primers based on the most similar sequences within 16 S rRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar ,...

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Bibliographic Details
Published in:World journal of microbiology & biotechnology 2022-07, Vol.38 (7), p.128-128, Article 128
Main Authors: Lai, Jyhmirn, Lin, Hengching, Hsu, Peichuan, Gondaira, Satoshi, Higuchi, Hidetoshi, Nagahata, Hajime
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Coagulase
Deoxyribonucleic acid
DNA
E coli
Environmental Engineering/Biotechnology
Klebsiella
Life Sciences
Mastitis
Microbiology
Mycoplasma
Original Paper
Pathogens
Polymerase chain reaction
Primers
rRNA
Sensitivity
Streptococcus infections
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Summary:The study aimed to develop a pair of polymerase chain reaction primers for detecting ruminant mycoplasma pathogens. We designed a set of primers based on the most similar sequences within 16 S rRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar , M. arginine , M. canadense , M. bovis , M. alkalescens , M. californicum , and M. bovigenitalium within the annealing temperature range of 46 to 48 °C. The minimum amount of DNA that can be detected using the protocol is 250 ng, which is equivalent to 2,000 colony-forming units per mL. The primers can detect mycoplasma from DNA extracted directly from milk samples. The common bovine mastitis pathogens of Staphylococcus aureus coagulase-negative staphylococci, Escherichia coli , Streptococcus uberis , Klebsiella pneumonia , and Kocuria rosea were not detected by the primers. We believe the high sensitivity and specificity of these primers make them useful for detecting infection with seven Mycoplasma species in ruminants, allowing the primers to be used in clinical settings.
ISSN:0959-3993
1573-0972
DOI:10.1007/s11274-022-03312-6