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Open Access Research article

Nasal swab samples and real-time polymerase chain reaction assays in community-based, longitudinal studies of respiratory viruses: the importance of sample integrity and quality control

Asma N Alsaleh123*, David M Whiley12, Seweryn Bialasiewicz12, Stephen B Lambert124, Robert S Ware15, Michael D Nissen126, Theo P Sloots126 and Keith Grimwood12

Author Affiliations

1 Queensland Children’s Medical Research Institute, The University of Queensland, Brisbane Queensland 4029, Australia

2 Queensland Paediatric Infectious Disease Laboratory, Royal Children’s Hospital, Brisbane Queensland 4029, Australia

3 Department of Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia

4 Queensland Health Communicable Diseases Branch, Queensland Health, Brisbane Queensland 4006, Australia

5 School of Population Health, The University of Queensland, Brisbane Queensland 4006, Australia

6 Microbiology Division, Pathology Queensland Central Laboratory, Brisbane Queensland 4006, Australia

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BMC Infectious Diseases 2014, 14:15  doi:10.1186/1471-2334-14-15

Published: 9 January 2014

Abstract

Background

Carefully conducted, community-based, longitudinal studies are required to gain further understanding of the nature and timing of respiratory viruses causing infections in the population. However, such studies pose unique challenges for field specimen collection, including as we have observed the appearance of mould in some nasal swab specimens. We therefore investigated the impact of sample collection quality and the presence of visible mould in samples upon respiratory virus detection by real-time polymerase chain reaction (PCR) assays.

Methods

Anterior nasal swab samples were collected from infants participating in an ongoing community-based, longitudinal, dynamic birth cohort study. The samples were first collected from each infant shortly after birth and weekly thereafter. They were then mailed to the laboratory where they were catalogued, stored at -80°C and later screened by PCR for 17 respiratory viruses. The quality of specimen collection was assessed by screening for human deoxyribonucleic acid (DNA) using endogenous retrovirus 3 (ERV3). The impact of ERV3 load upon respiratory virus detection and the impact of visible mould observed in a subset of swabs reaching the laboratory upon both ERV3 loads and respiratory virus detection was determined.

Results

In total, 4933 nasal swabs were received in the laboratory. ERV3 load in nasal swabs was associated with respiratory virus detection. Reduced respiratory virus detection (odds ratio 0.35; 95% confidence interval 0.27-0.44) was observed in samples where the ERV3 could not be identified. Mould was associated with increased time of samples reaching the laboratory and reduced ERV3 loads and respiratory virus detection.

Conclusion

Suboptimal sample collection and high levels of visible mould can impact negatively upon sample quality. Quality control measures, including monitoring human DNA loads using ERV3 as a marker for epithelial cell components in samples should be undertaken to optimize the validity of real-time PCR results for respiratory virus investigations in community-based studies.

Keywords:
Nasal swab; Respiratory virus; Real-time polymerase chain reaction; Quality control; Mould; Community-based study