Application of Alternative Nucleic Acid Extraction Protocols to ProGastro SSCS Assay for Detection of Bacterial Enteric Pathogens

Erik Munson; Maureen Napierala; Kimber L Munson; Dorothy Bilbo; Michael A Schulte

doi:10.1128/JCM.00271-16

Application of Alternative Nucleic Acid Extraction Protocols to ProGastro SSCS Assay for Detection of Bacterial Enteric Pathogens

J Clin Microbiol. 2016 May;54(5):1384-7. doi: 10.1128/JCM.00271-16. Epub 2016 Mar 2.

Authors

Erik Munson¹, Maureen Napierala², Kimber L Munson², Dorothy Bilbo², Michael A Schulte²

Affiliations

¹ College of Health Sciences, Marquette University, Milwaukee, Wisconsin, USA erik.munson@marquette.edu.
² Wheaton Franciscan Laboratory, Milwaukee, Wisconsin, USA.

Abstract

As an alternative to automated extraction, fecal specimens were processed by investigational lysis/heating (i.e., manual) and by chromatography/centrifugation (i.e., column) methods. ProGastro SSC and Shiga toxin-producing Escherichia coli (i.e., STEC) indeterminate rates for 101 specimens were 1.0% to 3.0% for automated, 11.9% for manual, and 24.8% to 37.6% for column methods. Following freeze-thaw of 247 specimens, indeterminate rates were 1.6% to 2.4% for manual and 0.8 to 5.3% for column methods. Mean processing times for manual and column methods were 30.5 and 69.2 min, respectively. Concordance of investigational methods with automated extraction was ≥98.8%.

Publication types

Evaluation Study

MeSH terms

Bacterial Infections / diagnosis*
Feces / microbiology*
Gastroenteritis / diagnosis*
Humans
Molecular Diagnostic Techniques / methods*
Nucleic Acids / isolation & purification*
Specimen Handling / methods*

Substances

Nucleic Acids