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

Alsaleh, Asma N., Whiley, David M., Bialasiewicz, Seweryn, Lambert, Stephen B., Ware, Robert S., Nissen, Michael D., Sloots, Theo P. and Grimwood, Keith (2014) 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. BMC Infectious Diseases, 14 1: . doi:10.1186/1471-2334-14-15


Author Alsaleh, Asma N.
Whiley, David M.
Bialasiewicz, Seweryn
Lambert, Stephen B.
Ware, Robert S.
Nissen, Michael D.
Sloots, Theo P.
Grimwood, Keith
Title 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
Journal name BMC Infectious Diseases   Check publisher's open access policy
ISSN 1471-2334
Publication date 2014-01-09
Year available 2014
Sub-type Article (original research)
DOI 10.1186/1471-2334-14-15
Open Access Status DOI
Volume 14
Issue 1
Total pages 9
Place of publication London, United Kingdom
Publisher BioMed
Language eng
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.
Formatted 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.
Keyword Nasal swab
Respiratory virus
Real-time polymerase chain reaction
Quality control
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID 615700
Institutional Status UQ
Additional Notes Article number 15.

 
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Created: Wed, 12 Feb 2014, 20:14:08 EST by Matthew Lamb on behalf of Child Health Research Centre