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Detecting laboratory DNA contamination using polyester-rayon wipes: a method validation study.

E Marielle Remillard¹, Laura K Taylor, Julie Layshock

¹Los Alamos National Laboratory, Los Alamos, NM 87545, USA.

Journal of Microbiological Methods

|January 16, 2013

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Laboratory surface testing using wipe methods can detect extraneous DNA, crucial for accurate PCR assays. However, a significant DNA loss occurs during the wipe wash step, impacting overall recovery efficiency.

Area of Science:

Molecular Biology
Laboratory Science

Background:

Polymerase chain reaction (PCR) assays are highly sensitive and prone to false positives from extraneous DNA.
Laboratory surface sampling using gauze wipes is a common method to detect environmental DNA contamination.

Purpose of the Study:

To evaluate the performance of a gauze wipe method for detecting extraneous DNA on laboratory surfaces.
To assess the accuracy, precision, limit of detection, linearity, and robustness of the wipe test method and its processing steps.

Main Methods:

The study utilized E. coli genomic DNA to quantify the performance of the wipe test method.
Key performance parameters including limit of detection and recovery efficiency were analyzed.
Robustness was tested through deliberate protocol modifications.

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Main Results:

The wipe test method achieved a limit of detection of 10^8 DNA copies, corresponding to 4.5x10^5 copies/area sampled.
A significant DNA loss (>58%) was observed during the wipe wash step, leading to a low recovery efficiency of 22±10%.
The method demonstrated robustness, maintaining consistent performance despite protocol variations.

Conclusions:

The gauze wipe method is robust for detecting extraneous DNA on laboratory surfaces.
Significant DNA loss during the wash step limits the method's accuracy and necessitates protocol optimization for improved recovery.