E-ISSN 2146-9369 | ISSN 2146-3158
 

Research Article 


Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels

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


Cited By:9

Abstract
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

Key words: 16S rRNA gene sequencing, Staphylococcus, Species-specific PCR


 
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How to Cite this Article
Pubmed Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. J Microbiol Infect Dis. 2018; 8(3): 120-127 . doi:10.5799/jmid.458462


Web Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. https://www.jmidonline.org/?mno=302656986 [Access: November 23, 2024]. doi:10.5799/jmid.458462


AMA (American Medical Association) Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. J Microbiol Infect Dis. 2018; 8(3): 120-127 . doi:10.5799/jmid.458462



Vancouver/ICMJE Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. J Microbiol Infect Dis. (2018), [cited November 23, 2024]; 8(3): 120-127 . doi:10.5799/jmid.458462



Harvard Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman (2018) Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. J Microbiol Infect Dis, 8 (3), 120-127 . doi:10.5799/jmid.458462



Turabian Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. 2018. Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. Journal of Microbiology and Infectious Diseases, 8 (3), 120-127 . doi:10.5799/jmid.458462



Chicago Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. "Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels." Journal of Microbiology and Infectious Diseases 8 (2018), 120-127 . doi:10.5799/jmid.458462



MLA (The Modern Language Association) Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman. "Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels." Journal of Microbiology and Infectious Diseases 8.3 (2018), 120-127 . Print. doi:10.5799/jmid.458462



APA (American Psychological Association) Style

Bibek Ranjan Shome, Krithiga Natesan, Susweta Das Mitra, Nimita Venugopal, Bhuvana Mani, Feroze Ganaie, Rajeswari Shome, Habibur Rehman (2018) Development of simplex-PCR assays for accurate identification of nine Staphylococcal species at genus and species levels. Journal of Microbiology and Infectious Diseases, 8 (3), 120-127 . doi:10.5799/jmid.458462