Medicine

Comparison of two multiplexed PCR assays for the detection of HSV-1,HSV-2, and VZV with extracted and unextracted cutaneous andmucosal specimens

Comparison of two multiplexed PCR assays for the detection of HSV-1,HSV-2, and VZV with extracted and unextracted cutaneous andmucosal specimens

Comparison of two multiplexed PCR assays for the detection of HSV-1,HSV-2, and VZV with extracted and unextracted cutaneous andmucosal specimens

Abstract

BACKGROUND:

Several analyte specific reagents (ASRs) are available for the detection and differentiation of HSV-1, HSV-2, and VZV in clinicalspecimens. However, there is limited data on the test performance of these reagents used in multiplexed PCR assays.

OBJECTIVE:

This study compared the performance of two multiplexed ASR sets for detection of HSV-1, HSV-2, and VZV in dermal specimens.

STUDY DESIGN:

Two commercially available ASRs were combined to produce multiplexed PCR assays for simultaneous detection of HSV-1, HSV-2, and VZV. Seeded samples were used to determine the limit of detection (LOD) for each assay. Patient samples (n=156) were tested in duplicate and results for each method compared to the reference standard of culture. Both extracted and unextracted specimens were used in the study.

RESULTS:

Both multiplexed PCR assays showed similar test performance, with minimal LOD differences observed. The LOD was 103copies/mL forHSV-1 and HSV-2 using the Focus assay compared to 5×103copies/mL and 2×104copies/mL, respectively for the EraGen assay. Both assaysshowed equal performance for VZV with a LOD of 5×103copies/mL. Analytical specificity testing showed no cross reactivity with other selected DNA viruses. Both assays showed similar performance when clinical samples were tested using both extracted and unextracted specimens.

CONCLUSION:

Commercially available ASRs can be successfully multiplexed for the PCR detection of HSV-1, HSV-2, and VZV using dermalspecimens. Either direct testing or nucleic acid extracted specimens can be used with similar performance in these assays.

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