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Molecular Detection of Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis

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Abstract

We developed molecular diagnostic assays for the detection of Streptococcus pyogenes (GAS) and Streptococcus dysgalactiae subsp. equisimilis (SDSE), two streptococcal pathogens known to cause both pharyngitis and more invasive forms of disease in humans. Two real-time PCR assays coupled with an internal control were designed to be performed in parallel. One assay utilizes a gene target specific to GAS, and the other utilizes a gene target common to the two species. Both assays showed 2–3 orders of magnitude improved analytical sensitivity when compared to a commercially available rapid antigen test. In addition, when compared to standard culture in an analysis of 96 throat swabs, the real-time PCR assays resulted in clinical sensitivity and specificity of 91.7 and 100%, respectively. As capital equipment costs for real-time PCR can be prohibitive in smaller laboratories, the real-time PCR assays were converted to a low-density microarray format designed to function with an inexpensive photopolymerization-based non-enzymatic signal amplification (NESA™) method. S. pyogenes was successfully detected on the low-density microarray in less than 4 h from sample extraction through detection.

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Acknowledgments

This work was supported in part by National Institutes of Health/National Institute of Allergy and Infectious Diseases (NIH/NIAID, grant number U01A1070276). The authors would like to acknowledge the Laboratory Services Division of the Colorado Department of Public Health and Environment (CDPHE) for providing culture plates, Boulder Community Hospital (BCH) for providing clinical specimens, associated culture results and matched culture plates, and Quidel Corporation for graciously providing the QuickVue® Dipstick Strep A rapid antigen tests.

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  1. James A. Smagala

    Present address: Influenza Division, The Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30333, USA

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  1. InDevR Inc., 2100 Central Avenue, Suite 106, Boulder, CO, 80301, USA

    Erica D. Dawson, Amber W. Taylor, James A. Smagala & Kathy L. Rowlen

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Correspondence to Erica D. Dawson.

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Dawson, E.D., Taylor, A.W., Smagala, J.A. et al. Molecular Detection of Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis . Mol Biotechnol 42, 117–127 (2009). https://doi.org/10.1007/s12033-009-9143-2

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