Dataset Information

Pitfalls in interpretation of CT-values of RT-PCR in children with acute respiratory tract infections.

ABSTRACT:

Background

The relation between viral load and disease severity in childhood acute respiratory tract infections (ARI) is not fully understood.

Objectives

To assess the clinical relevance of the relation between viral load, determined by cycle threshold (CT) value of real-time reverse transcription-polymerase chain reaction assays and disease severity in children with single- and multiple viral ARI.

Study design

582 children with ARI were prospectively followed and tested for 15 viruses. Correlations were calculated between CT values and clinical parameters.

Results

In single viral ARI, statistically significant correlations were found between viral loads of Respiratory Syncytial Virus (RSV) and hospitalization and between viral loads of Human Coronavirus (HCoV) and a disease severity score. In multiple-viral ARI, statistically significant correlations between viral load and clinical parameters were found. In RSV-Rhinovirus (RV) multiple infections, a low viral load of RV was correlated with a high length of hospital stay and a high duration of extra oxygen use. The mean CT value for RV, HCoV and Parainfluenza virus was significantly lower in single- versus multiple infections.

Conclusion

Although correlations between CT values and clinical parameters in patients with single and multiple viral infection were found, the clinical importance of these findings is limited because individual differences in host-, viral and laboratory factors complicate the interpretation of statistically significant findings. In multiple infections, viral load cannot be used to differentiate between disease causing virus and innocent bystanders.

SUBMITTER: Wishaupt JO

PROVIDER: S-EPMC7185604 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Publications

Pitfalls in interpretation of CT-values of RT-PCR in children with acute respiratory tract infections.

Wishaupt Jérôme O JO Ploeg Tjeerd van der TV Smeets Leo C LC Groot Ronald de R Versteegh Florens G A FG Hartwig Nico G NG

Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology 20170224

<h4>Background</h4>The relation between viral load and disease severity in childhood acute respiratory tract infections (ARI) is not fully understood.<h4>Objectives</h4>To assess the clinical relevance of the relation between viral load, determined by cycle threshold (CT) value of real-time reverse transcription-polymerase chain reaction assays and disease severity in children with single- and multiple viral ARI.<h4>Study design</h4>582 children with ARI were prospectively followed and tested fo ...[more]

PMID: 28259567

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Project description:Coronavirus disease 2019 (COVID-19) is an infectious, acute respiratory disease caused mainly by person-to-person transmission of the coronavirus SARS-CoV-2. Its emergence has caused a world-wide acute health crisis, intensified by the challenge of reliably identifying individuals likely to transmit the disease. Diagnosis is hampered by the many unknowns surrounding this disease, including those relating to infectious viral burden. This uncertainty is exacerbated by disagreement surrounding the clinical relevance of molecular testing using reverse transcription quantitative PCR (RT-qPCR) for the presence of viral RNA, most often based on the reporting of quantification cycles (Cq), which is also termed the cycle threshold (Ct) or crossing point (Cp). Despite it being common knowledge that Cqs are relative values varying according to a wide range of different parameters, there have been efforts to use them as though they were absolute units, with Cqs below an arbitrarily determined value, deemed to signify a positive result and those above, a negative one. Our results investigated the effects of a range of common variables on Cq values. These data include a detailed analysis of the effect of different carrier molecules on RNA extraction. The impact of sample matrix of buccal swabs and saliva on RNA extraction efficiency was demonstrated in RT-qPCR and the impact of potentially inhibiting compounds in urine along with bile salts were investigated in RT-digital PCR (RT-dPCR). The latter studies were performed such that the impact on the RT step could be separated from the PCR step. In this way, the RT was shown to be more susceptible to inhibitors than the PCR. Together, these studies demonstrate that the consequent variability of test results makes subjective Cq cut-off values unsuitable for the identification of infectious individuals. We also discuss the importance of using reliable control materials for accurate quantification and highlight the substantial role played by dPCR as a method for their development.

Project description:ObjectiveTo determine the expected duration of symptoms of common respiratory tract infections in children in primary and emergency care.DesignSystematic review of existing literature to determine durations of symptoms of earache, sore throat, cough (including acute cough, bronchiolitis, and croup), and common cold in children.Data sourcesPubMed, DARE, and CINAHL (all to July 2012).Eligibility criteria for selecting studiesRandomised controlled trials or observational studies of children with acute respiratory tract infections in primary care or emergency settings in high income countries who received either a control treatment or a placebo or over-the-counter treatment. Study quality was assessed with the Cochrane risk of bias framework for randomised controlled trials, and the critical appraisal skills programme framework for observational studies.Main outcome measuresIndividual study data and, when possible, pooled daily mean proportions and 95% confidence intervals for symptom duration. Symptom duration (in days) at which each symptom had resolved in 50% and 90% of children.ResultsOf 22,182 identified references, 23 trials and 25 observational studies met inclusion criteria. Study populations varied in age and duration of symptoms before study onset. In 90% of children, earache was resolved by seven to eight days, sore throat between two and seven days, croup by two days, bronchiolitis by 21 days, acute cough by 25 days, common cold by 15 days, and non-specific respiratory tract infections symptoms by 16 days.ConclusionsThe durations of earache and common colds are considerably longer than current guidance given to parents in the United Kingdom and the United States; for other symptoms such as sore throat, acute cough, bronchiolitis, and croup the current guidance is consistent with our findings. Updating current guidelines with new evidence will help support parents and clinicians in evidence based decision making for children with respiratory tract infections.

Dataset Information

Pitfalls in interpretation of CT-values of RT-PCR in children with acute respiratory tract infections.

Background

Objectives

Study design

Results

Conclusion

Publications

Pitfalls in interpretation of CT-values of RT-PCR in children with acute respiratory tract infections.

Similar Datasets

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