A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
PCR inhibition in stool samples in relation to age of infants
Tekijät: Oikarinen S, Tauriainen S, Viskari H, Simell O, Knip M, Virtanen S, Hyoty H
Kustantaja: ELSEVIER SCIENCE BV
Julkaisuvuosi: 2009
Journal: Journal of Clinical Virology
Tietokannassa oleva lehden nimi: JOURNAL OF CLINICAL VIROLOGY
Lehden akronyymi: J CLIN VIROL
Vuosikerta: 44
Numero: 3
Aloitussivu: 211
Lopetussivu: 214
Sivujen määrä: 4
ISSN: 1386-6532
DOI: https://doi.org/10.1016/j.jcv.2008.12.017
Tiivistelmä
Background: PCR is rapidly replacing traditional methods in diagnostic virus laboratories. PCR inhibitors,
which are often present in clinical samples, may lead to false negative test results.
Objectives: The aim was to study the presence of PCR inhibitors in stool samples collected from 3- to
24-month old children.
Study design: Total RNA fraction extracted from stool samples was spiked with a standardized amount of
Semliki Forest Virus RNA and amplified using specific PCR primers. The presence of PCR inhibitors was
detected by a decrease in amplification rate compared to spiked water samples. Inhibition in different
age groups and dietary origin of PCR inhibitors were analyzed by comparing the samples taken during
exclusive and non-exclusive breastfeeding periods. The inactivation of PCR inhibitors was also assessed.
Results: Complete inhibition was seen in 12% (13/108) and partial inhibition in 19% (21/108) of the samples.
Inhibition was seen in none of the stool samples (0/31) taken from infants younger than 6 months
compared to 17% of samples (13/77) taken from6 to 24 months old infants (p < 0.036). Breastfeeding was
more common in younger age group. Addition of bovine serum albumin (BSA) into the reaction mixtures
eliminated the effect of inhibitors leading to all samples being positive.
Conclusions: PCR inhibitors are frequent in stool samples. They may originate from dietary components
and can lead to false negative PCR results. The addition of BSA to the cDNA and PCR reactions proved to
be an easy and effective method for eliminating the inhibitory effect of these compounds.
Background: PCR is rapidly replacing traditional methods in diagnostic virus laboratories. PCR inhibitors,
which are often present in clinical samples, may lead to false negative test results.
Objectives: The aim was to study the presence of PCR inhibitors in stool samples collected from 3- to
24-month old children.
Study design: Total RNA fraction extracted from stool samples was spiked with a standardized amount of
Semliki Forest Virus RNA and amplified using specific PCR primers. The presence of PCR inhibitors was
detected by a decrease in amplification rate compared to spiked water samples. Inhibition in different
age groups and dietary origin of PCR inhibitors were analyzed by comparing the samples taken during
exclusive and non-exclusive breastfeeding periods. The inactivation of PCR inhibitors was also assessed.
Results: Complete inhibition was seen in 12% (13/108) and partial inhibition in 19% (21/108) of the samples.
Inhibition was seen in none of the stool samples (0/31) taken from infants younger than 6 months
compared to 17% of samples (13/77) taken from6 to 24 months old infants (p < 0.036). Breastfeeding was
more common in younger age group. Addition of bovine serum albumin (BSA) into the reaction mixtures
eliminated the effect of inhibitors leading to all samples being positive.
Conclusions: PCR inhibitors are frequent in stool samples. They may originate from dietary components
and can lead to false negative PCR results. The addition of BSA to the cDNA and PCR reactions proved to
be an easy and effective method for eliminating the inhibitory effect of these compounds.
Ladattava julkaisu This is an electronic reprint of the original article. |  |
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