Real-time PCR assay for direct quantification of Staphylococcus aureus DNA in blood samples

2016 ◽  
Author(s):  
Luis Eduardo Lopez-Cortes
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Assay ◽  
Blood Samples ◽  
Direct Quantification

scholarly journals Detection of Aspergillus species in bone marrow transplant patients

2010 ◽  
Vol 4 (08) ◽  
pp. 511-516 ◽  
Author(s):  
Parisa Badiee ◽  
Abdolvahab Alborzi
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Clinical Signs ◽  
Marrow Transplant ◽  
Aspergillus Species ◽  
Pcr Assay ◽  
Blood Samples ◽  
Transplant Patients

Introduction:  Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation.  The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.

scholarly journals Quantification by Real-Time PCR Assay of Staphylococcus aureus Load: a Useful Tool for Rapidly Identifying Persistent Nasal Carriers

2012 ◽  
Vol 50 (6) ◽  
pp. 2063-2065 ◽  
Author(s):  
Paul O. Verhoeven ◽  
Florence Grattard ◽  
Anne Carricajo ◽  
Frédéric Lucht ◽  
Céline Cazorla ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Assay

scholarly journals Real-Time PCR Assay for Detection of blaZ Genes in Staphylococcus aureus Clinical Isolates

2014 ◽  
Vol 52 (4) ◽  
pp. 1259-1261 ◽  
Author(s):  
L. A. Pereira ◽  
G. B. Harnett ◽  
M. M. Hodge ◽  
J. A. Cattell ◽  
D. J. Speers
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Pcr Assay

scholarly journals Real-Time PCR Assay for Detection of Fluoroquinolone Resistance Associated with grlA Mutations in Staphylococcus aureus

2003 ◽  
Vol 41 (7) ◽  
pp. 3246-3251 ◽  
Author(s):  
P. Lapierre ◽  
A. Huletsky ◽  
V. Fortin ◽  
F. J. Picard ◽  
P. H. Roy ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Fluoroquinolone Resistance ◽  
Pcr Assay

scholarly journals Real-time evaluation of an optimized real-time PCR assay versus Brilliance chromogenic MRSA agar for the detection of meticillin-resistant Staphylococcus aureus from clinical specimens

2011 ◽  
Vol 60 (3) ◽  
pp. 323-328 ◽  
Author(s):  
J. Danial ◽  
M. Noel ◽  
K. E. Templeton ◽  
F. Cameron ◽  
F. Mathewson ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Test Performance ◽  
High Sensitivity ◽  
Turnaround Time ◽  
Homology Search ◽  
Pcr Assay ◽  
Indirect Measure ◽  
The Mean

A total of 1204 meticillin-resistant Staphylococcus aureus (MRSA) screens (3340 individual swabs) were tested to evaluate a staphylococcal cassette chromosome mec (SCCmec) real-time PCR. In total, 148 (12.3 %) of the screens were MRSA-positive, where 146 (12.1 %) were MRSA-positive by the SCCmec real-time PCR assay. In contrast, 128 (10.6 %) screens were MRSA-positive by culture. One hundred and twenty-six (10.5 %) of the screens were positive by both culture and PCR. Twenty of the 1204 screens (1.66 %) were negative by culture but positive by PCR; these samples were sequenced. In 14 of the cases, a homology search confirmed the sequence as SCCmec, indicating that these samples could be considered true positives. Two of the 1204 (0.2 %) screens were positive by culture and negative by PCR. The mean turnaround time (TAT) for PCR-negative swabs was 6 h 12 min and for PCR-positive swabs was 6 h 48 min. In comparison, for culture-negative swabs the mean TAT was 29 h 30 min and for culture-positive swabs was 69 h. The cost per swab for routine culture was £0.41 (€0.48) and that of the real-time PCR assay was £2.35 (€2.75). This optimized, in-house, inexpensive, real-time PCR test maintained a very high sensitivity and specificity when evaluated under real-time laboratory conditions. The TAT of this real-time PCR assay was substantially lower than that of chromogenic culture. It was also maintained throughout the entire process, which can be taken as an indirect measure of test performance. This study showed that implementation of a molecular test can be achieved with limited resources in a standard microbiology laboratory.

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