Rapid Detection of the Staphylococcal mecA Gene from BACTEC Blood Culture Bottles by the Polymerase Chain Reaction (original) (raw)

Journal Article

,

1 From the Associated Regional and University Pathologists, Inc., 500 Chipeta Way, Salt Lake City, Utah

2 From the Deparlment of Pathology, University of Utah Medical Center, 50 N. Medical Drive. Salt Lake City, Utah.

Search for other works by this author on:

,

1 From the Associated Regional and University Pathologists, Inc., 500 Chipeta Way, Salt Lake City, Utah

Search for other works by this author on:

,

Patricia L. Newcomb-Gayman, MT(ASCP)

1 From the Associated Regional and University Pathologists, Inc., 500 Chipeta Way, Salt Lake City, Utah

Search for other works by this author on:

1 From the Associated Regional and University Pathologists, Inc., 500 Chipeta Way, Salt Lake City, Utah

2 From the Deparlment of Pathology, University of Utah Medical Center, 50 N. Medical Drive. Salt Lake City, Utah.

Search for other works by this author on:

Published:

01 November 1996

• PDF
• Split View
• • Article contents
  • Figures & tables
  • Video
  • Audio
  • Supplementary Data
• Cite

Cite

Karen C. Carroll, Rebecca B. Leonard, Patricia L. Newcomb-Gayman, David R. Hillyard, Rapid Detection of the Staphylococcal _mec_A Gene from BACTEC Blood Culture Bottles by the Polymerase Chain Reaction, American Journal of Clinical Pathology, Volume 106, Issue 5, 1 November 1996, Pages 600–605, https://doi.org/10.1093/ajcp/106.5.600
Close

Navbar Search Filter Mobile Enter search term Search

Abstract

A rapid polymerase chain reaction (PCR) method for the direct detection of the staphylococcal mecA gene from BACTEC blood culture bottles (Becton Dickinson, Sparks, MD) was developed. Published primer sequences and sample preparation using Achromopeptidase for cell lysis were adapted to the use of the Idaho Technology Air Thermocycler 1605 (Idaho Technologies, Idaho Falls, ID).

The method was validated with 80 strains of coagulase-positive and coagulase-negative geographically diverse methicillin-resistant and susceptible isolates of staphylococci. There was a 100% correlation between the PCR results and the results of standard susceptibility testing methods.

From BACTEC 9240 blood cultures, mixed aliquots of blood and broth containing gram-positive cocci in clusters were centrifuged at low speed to sediment red blood cells. After additional centrifugation and wash steps, PCR was performed on the resuspended pellet. The turnaround time from initial Gram stain detection of positive BACTEC bottles to PCR amplicon detection by agarose gel electrophoresis is less than 3 hours.

In a clinical evaluation of 181 blood culture isolates, there was a 99% correlation with standard susceptibility results for Staphylococcus aureus. Discrepant results for Staphylococcus aureus isolates were verified by a Mueller Hinton plate supplemented with 6 μg/mL of oxacillin and 2% sodium chloride. For coagulase-negative staphylococci, the PCR method detected an additional seven resistant isolates that were reported by the Vitek as susceptible. Coagulase-negative staphylococcal susceptibility results that were in disagreement with the PCR assay were confirmed by the disk-diffusion method.

This procedure is accurate, rapid and fits well into laboratory work flow. Rapid detection of the mecA gene on positive blood culture vials has become a routine test in the authors’ clinical microbiology laboratory.

This content is only available as a PDF.

© American Society for Clinical Pathology

Citations

Views

Altmetric

Metrics

Total Views 588

54 Pageviews

534 PDF Downloads

Since 12/1/2016

×

Email alerts

Citing articles via

• Latest

• Most Read

• Most Cited

More from Oxford Academic