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Background <p>Buccal swabs are widely used as a non-invasive method for genomic DNA collection in large-scale genotyping studies. However, the sporadic presence of PCR inhibitors within these samples may hinder PCR amplification and affect assay reliability. This study investigates the incorporation of bovine serum albumin (BSA) into the PCR reaction mixture as an additive to counteract PCR inhibition.</p> Results <p>In our high-throughput setting, the incorporation of BSA significantly improved robustness, lowering failure rates to 0.1% in subsequent routine operation across 1,000,000 buccal swab samples. Despite minor challenges with foaming during automated liquid handling, no detrimental effects on PCR performance were observed.</p> Conclusion <p>These results underscore the efficacy of BSA in improving PCR robustness, enhancing the reliability of high-throughput molecular diagnostic assays.</p>

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PCR optimization for buccal swab-derived samples: overcoming sporadic inhibition with bovine serum albumin

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年11月21日

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  • Stefanie Tietze,
  • Julia Phieler,
  • Sabine Bergmann,
  • Juliane Grimmer,
  • Kathrin Putke,
  • Bastian Joffroy,
  • Yannic Utz,
  • Katrin Hengst,
  • Bianca Heyn,
  • Diana Panhans,
  • Alexander H. Schmidt,
  • Anja Klussmeier,
  • Vinzenz Lange

摘要

Background

Buccal swabs are widely used as a non-invasive method for genomic DNA collection in large-scale genotyping studies. However, the sporadic presence of PCR inhibitors within these samples may hinder PCR amplification and affect assay reliability. This study investigates the incorporation of bovine serum albumin (BSA) into the PCR reaction mixture as an additive to counteract PCR inhibition.

Results

In our high-throughput setting, the incorporation of BSA significantly improved robustness, lowering failure rates to 0.1% in subsequent routine operation across 1,000,000 buccal swab samples. Despite minor challenges with foaming during automated liquid handling, no detrimental effects on PCR performance were observed.

Conclusion

These results underscore the efficacy of BSA in improving PCR robustness, enhancing the reliability of high-throughput molecular diagnostic assays.