RRML - Protocol and Matters for Consideration for the Treatment of Polymerase Chain Reaction Contamination in Next-Generation Sequencing Laboratories
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Nr. 30(4)/2022 DOI:10.2478/rrlm-2022-0038
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Original professional paper

Protocol and Matters for Consideration for the Treatment of Polymerase Chain Reaction Contamination in Next-Generation Sequencing Laboratories

Lin Wang, Chen Ma, Bin He, Rong Qiang, Hongsu Zhang

Correspondence should be addressed to: Lin Wang

Abstract:

Objective: The contamination of polymerase chain reaction (PCR) samples in molecular diagnostic laboratories can cause serious consequences. Internal quality control efforts are often inadequate, especially in clinical next-generation sequencing (NGS) laboratories. Methods: In this study, we retrospectively investigated an incidence of PCR contamination and its decontamination process in a clinical laboratory. We performed a series of measures for decontamination. Taqman fluorescence quantification was carried out to determine the presence of contaminating DNA. SYBR-Green PCR was conducted to evaluate the effect of chlorine disinfectant on NGS library preparation. Results: Through a series of elimination measures undertaken over 8 weeks, the decontamination process was verified as reliable. Almost no contamination was detected. Chlorine disinfectant should be forbidden in Illumina NGS laboratories because it may cause the failure of library preparation.Conclusion: Our prevention and decontamination strategies could effectively eliminate PCR amplicons. Chlorine disinfectants should not be used in Illumina NGS laboratories.

Keywords: laboratory contamination; chlorine disinfectants; high-throughput library

Received: 15.6.2022
Accepted: 14.9.2022
Published: 30.9.2022

 
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How to cite
Wang L, Ma C, He B, Qiang R, Zhang H. Protocol and Matters for Consideration for the Treatment of Polymerase Chain Reaction Contamination in Next-Generation Sequencing Laboratories. Rev Romana Med Lab. 2022;30(4):467-75. DOI:10.2478/rrlm-2022-0038