Journal List > Lab Med Online > v.3(4) > 1057210

Lee, Park, Yoon, Lee, and Cho: Evaluation of ExiPrep16 Automated System for the Extraction of Nucleic Acids from Nasopharyngeal Swabs for the Detection of Respiratory Viruses

Abstract

Background

Automated nucleic acid extraction offers a standardized sample treatment method, low error rate, and avoids sample nucleic acid contamination for use in molecular diagnostics. Here, we evaluated the performance of automated ExiPrep16 system (Bioneer Co.) in comparison with the manual Viral Gene-spin Viral DNA/RNA Extraction kit (VGspin; iNtRON Biotechnology Inc.) for the detection of respiratory viruses from nasopharyngeal flocked swabs.

Methods

To compare the agreement rate and analytical sensitivity between ExiPrep16 and VGspin, previously collected 78 patient samples and 11 pooled samples of each respiratory viruses and their serially diluted samples (until 1/108), were tested by multiplex reverse-transcriptase PCR (Seeplex RV 12 ACE Detection kit; SeeGene Inc.). In addition, we repeatedly analyzed the threshold cycle of the pooled and 1/103 dilution of adenovirus (ADV) and influenza virus A (Flu-A) by using real-time PCR to evaluate the precision and crossover of the ExiPrep16 system.

Results

The analytical sensitivity of the ExiPrep16 was comparable to that of VGspin, and the highest detectable dilution varied in the range of 1/10 to 1/106 depending on the viruses. The total, overall positive and negative percent agreements of ExiPrep16 in comparison with VGspin were 95.7%, 96.2%, and 95.2%, respectively. The mean (CV%) of pooled and 1/103 dilution of ADV were, respectively, 19.2 cycle (2.1%) and 31.6 cycle (4.3%) and those for Flu-A were 22.6 cycle (3.1%) and 35.5 cycle (2.6%). No carryover was detected.

Conclusions

Compared to the manual VGspin, ExiPrep16 ensured nucleic acid extraction for efficient detection of respiratory viruses.

Figures and Tables

Table 1
Comparison of the analytical sensitivity between manual Viral gene-spin Viral DNA/RNA Extraction kit (VGspin) and automatic ExiPrep16 system
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+: thick band, w+: thin band confirmed repeatedly, ww+: thin band, but cannot be confirmed repeatedly.

Abbreviations: ADV, adenovirus; Flu-A, influenza virus A; Flu-B, influenza virus B; PIV, parainfluenza virus; RSV, respiratory syncytial virus; hMPV, human metapneumovirus; hRV, human rhinovirus.

Table 2
Agreement rate between manual VGspin kitand automatic ExiPrep16 system for the detection of respiratory viruses in flocked nasopharyngeal swabs
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*Measurement of agreement (K) between ExiPrep16 and VGspin was 0.914; including 1 case of RSV-A and 1 case of hRV-A; including 1 case of hMPV and 1 case of ADV.

Table 3
Reproducibility of Ct values analysed by real-time PCR using nucleic acid extracted by automatic ExiPrep16 system
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Abbreviations: Ct, threshold cycle; CI, confidence interval.

Notes

This article is available from http://www.labmedonline.org

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