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Performance evaluation of an automated magnetic bead based DNA extraction and real-time PCR for detection of BK virus
  • +5
  • Ying Gu,
  • Xiaodong Fan,
  • Yuting He,
  • Hao Huang,
  • Xuegao Yu,
  • Gang Huang,
  • Huang Bin,
  • Peisong Chen
Ying Gu
Shenzhen Longhua New District People's Hospital
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Xiaodong Fan
The People’s Hospital of Wenshan Prefecture
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Yuting He
Sun Yat-sen University First Affiliated Hospital Department of Laboratory Medicine
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Hao Huang
Sun Yat-sen University First Affiliated Hospital
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Xuegao Yu
Sun Yat-sen University First Affiliated Hospital
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Gang Huang
Sun Yat-sen University First Affiliated Hospital
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Huang Bin
Sun Yat-sen University First Affiliated Hospital
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Peisong Chen
Sun Yat-sen University First Affiliated Hospital

Corresponding Author:[email protected]

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Abstract

Background: To evaluate the analytical performance of a magnetic bead-based automatic extraction platform DAAN Stream SP96 combined with quantitative real-time PCR (qPCR) in detection of BK virus. Methods: A total of 70 clinical specimens from renal transplant recipients (RTR) were analyzed. The evaluation was carried out according to CLSI EP documents. The performance parameters included precision, accuracy, linearity, detection limit, interference test, cross-reactivity, and method comparison. Results: The coefficient of variation (CV) for within-run and intermediate precision of the high-, middle-, and low-level samples were all <5%. For the accuracy, the mean percent bias was 0.6% compared with the reference method. The linear range was 5.0 × 103~5.0 × 108 copies/mL, and the limit of detection (LOD) was 1.23×103 copies/mL. For anti-interference ability, 280 μmol/L bilirubin, 18mmol /L triglycerides, and 20g /L hemoglobin had no apparent effects on testing results. This assay didn’t show cross-reaction with some common pathogenic viruses. The automated magnetic bead-based DNA extraction combined qPCR has a good correlation (R2= 0.8978) with the manual boiling extraction combined qPCR. Conclusion: The DAAN Stream SP96 magnetic bead-based automatic extraction platform combined qPCR showed good analytical performance in the BKV DNA quantitative detection, which can meet the clinical testing requirements well.