Detection of methicillin- and aminoglycoside-resistant genes and simultaneous identification of S. aureus using triplex real-time PCR Taqman assay

Sabet, Negar Shafiei; Subramaniam, G.; Parasakthi, N; Sekaran, Shamala Devi

doi:10.1016/j.mimet.2006.07.008

Cited by 22 publications

(9 citation statements)

References 23 publications

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“…S. aureus identity and meticillin resistance were confirmed by PCR amplification of the coagulase gene and mecA gene, as described elsewhere (Sabet et al, 2007). We also tested for the presence of the PVL gene by PCR according to a previously described method (McClure et al, 2006).…”

Section: Methodsmentioning

confidence: 99%

Population structure of colonizing and invasive Staphylococcus aureus strains in northern Vietnam

Jafari

Aardema

et al. 2016

Journal of Medical Microbiology

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Staphylococcus aureus is an important global health problem worldwide. There is still scarce information on the population structure of S. aureus strains in Asia, where the majority of the world population lives. This study characterized the diversity of S. aureus strains in northern Vietnam through multilocus sequence typing (MLST). Eighty-five carriage isolates from the community and 77 invasive isolates from the clinical setting were selected and tested for meticillin resistance and the presence of Panton-Valentine leukocidin (PVL). MLST was performed on these isolates, of which CC59 (25.4 %), CC188 (17.3 %) and CC45 (16.7 %) were the predominant clonal complexes (CCs). CC59 carriage isolates had significantly lower rates of meticillin-resistant S. aureus (MRSA) than their corresponding clinical group isolates (32 vs 83 %). There were no significant differences in rates of MRSA between carriage isolates and clinical isolates of CC45 and CC188. CC59 carriage isolates were significantly lower in rates of PVL + than CC59 clinical isolates (32 vs 83 %), but the converse was shown in CC45isolates (14 vs 0 %, respectively). This study revealed vast differences in the molecular epidemiology and population structure of S. aureus in community and clinical settings in Vietnam. Nevertheless, the data underline the spread of virulent and/or resistant strains (MRSA and/or PVL + ) in the community, suggesting the necessity for further surveillance to determine the mechanism of transmission of these strains (i.e. MRSA/PVL + ) outside clinical settings.

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Section: Methodsmentioning

confidence: 99%

Population structure of colonizing and invasive Staphylococcus aureus strains in northern Vietnam

Jafari

Aardema

et al. 2016

Journal of Medical Microbiology

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“…Furthermore, real-time PCR is well suited for screening a large number of samples at the same time; that allows a rapid, reliable, efficient and less costly routine laboratory diagnosis (Kunori et al 2002;Tan et al 2001). The benefit of accuracy and speed in simultaneous identification of species and detection of methicillin-susceptibility could make this method a valuable diagnostic tool, especially in hospitals in areas where MRSA is endemic (Sabet et al 2007;Grisold et al 2002).…”

Section: Discussionmentioning

confidence: 99%

“…However, this type of PCR assay still quite laborious and time-consuming since it requires the preparation and PCR product characterization using gel electrophoresis (Costa et al 2005;Kearns et al 1999). Recently, a number of real-time PCR studies based on fluorescent probes have been carried out to detect MRSA from various clinical sources using TaqMan probes (Sabet et al 2006(Sabet et al , 2007Costa et al 2005). However, these assays require the availability of primers and probes that must be selected according to very rigid conditions, which cannot always be easily applied (Nam et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Detection of Malaysian methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates using simplex and duplex real-time PCR

Suhaili

Johari

Mohtar

et al. 2008

World J Microbiol Biotechnol

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The aim of this study was to develop a methicillin-resistant Staphylococcus aureus (MRSA) detection method based on the melting temperature analysis profiling of S. aureus clinical isolates from three different hospitals in Malaysia. Simplex and duplex real-time PCR assay was used for the simultaneous detection of nuc (species-specific) and mecA (methicillin-resistance) genes in a single SYBR Green I real-time PCR tube assay. Evaluations were based on the melting temperature (T m ) analysis of the amplicons using 23 S. aureus clinical isolates including three ATCC S. aureus standard strains. Real-time PCR amplification products with melting peaks at 78.39 ± 0.4°C and 74.41 ± 0.6°C were detected for nuc and mecA genes, respectively. Each real-time PCR assay was completed within two hours. This rapid genotypic method is useful for the detection of resistant determinant (mecA) and identification of S. aureus (nuc) clinical isolates, thus benefiting patient therapy in hospitals.

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“…12,13 Although PCR is a highly sensitive technique that can be used in direct identiÞ cation of bacteria in blood, 12,13 it may not be appropriate for daily routine work as it is time-consuming, expensive and expertise demanding compared to FISH, which is rapid and inexpensive. Furthermore, the added advantage of FISH over PCR is that extraction of DNA from bacteria is not required in the former.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of non-enterobacteriaceae directly from positive blood culture using fluorescent <i>In situ </i>hybridization

Wong¹,

Subramaniam²,

Parasakthi³

et al. 2007

Indian J Med Microbiol

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Detection of methicillin- and aminoglycoside-resistant genes and simultaneous identification of S. aureus using triplex real-time PCR Taqman assay

Cited by 22 publications

References 23 publications

Population structure of colonizing and invasive Staphylococcus aureus strains in northern Vietnam

Population structure of colonizing and invasive Staphylococcus aureus strains in northern Vietnam

Detection of Malaysian methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates using simplex and duplex real-time PCR

Rapid detection of non-enterobacteriaceae directly from positive blood culture using fluorescent <i>In situ </i>hybridization

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