Development of Simplex-PCR assays for Accurate Identification of Nine Staphylococcal Species at Genus and Species Levels

Yıl 2018, Cilt: 08 Sayı: 03, 120 - 127, 15.09.2018

Bibek Ranjan Shome Krithiga Natesan Susweta Das Mitra Nimita Venugopal Bhuvana Mani Feroze Ganaie Rajeswari Shome Habibur Rahman

https://doi.org/10.5799/jmid.458462

Cited By: 2

Öz

Objective: Species identification of Staphylococcus is the prerequisite for precise assessment of
microbial dynamics including their transmission and pathogenic significance in
the dairy herd environment. The present study aimed to develop simplex PCR
assays for rapid and specific identification of nine different Staphylococcus species.

Methods: Specific
primers targeting sodA gene for S. aureus, S. chromogenes, S. hominis,
S. haemolyticus, S. hyicus; gap gene for S. sciuri, S. auricularis, S. simulans;
and rdr gene for S. epidermidis were designed. The PCR assays were evaluated against
28 ATCC reference strains and 209 composite milk samples. Partial 16S rRNA sequencing was performed to
reconfirm the results.

Results: The
PCR assays allowed species level identification of 348 staphylococcal field
isolates recovered from 209 milk samples. The identification pattern was S. aureus (n=101), S. chromogenes (n=89), S.
epidermidis (n=57), S. sciuri
(n=43), S. haemolyticus (n=34), S. hyicus (n=13), S. hominis (n=5), S.
auricularis (n=3) and S. simulans (n=3). The PCR based species
identification was in 100% concordance with the partial 16S rRNA gene sequencing approach.

Conclusion: The simplex PCR assays can be used as a precise tool
for routine identification of Staphylococcus
species from bovine milk as Staphylococcus
species including coagulase-negative staphylococci is recognized a major cause
of bovine mastitis in different parts of the world including India. J Microbiol Infect Dis 2018; 8(3):120-127

Anahtar Kelimeler

16S rRNA gene sequencing, Staphylococcus, Species-specific PCR

Kaynakça

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