Abstract

BackgroundChlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest. A number of different PCR assays have been developed and some are used in combination with serology for diagnosis. Real-time PCR could be an attractive new PCR method; therefore it must be evaluated and compared to conventional PCR methods.ResultsWe compared the performance of a newly developed real-time PCR with a conventional PCR method for detection of C. pneumoniae. The PCR methods were tested on reference samples containing C. pneumoniae DNA and on 136 nasopharyngeal samples from patients with a chronic cough. We found the same detection limit for the two methods and that clinical performance was equal for the real-time PCR and for the conventional PCR method, although only three samples tested positive. To investigate whether the low prevalence of C. pneumoniae among patients with a chronic cough was caused by suboptimal PCR efficiency in the samples, PCR efficiency was determined based on the real-time PCR. Seventeen of twenty randomly selected clinical samples had a similar PCR efficiency to samples containing pure genomic C. pneumoniae DNA.ConclusionsThese results indicate that the performance of real-time PCR is comparable to that of conventional PCR, but that needs to be confirmed further. Real-time PCR can be used to investigate the PCR efficiency which gives a rough estimate of how well the real-time PCR assay works in a specific sample type. Suboptimal PCR efficiency of PCR is not a likely explanation for the low positivity rate of C. pneumoniae in patients with a chronic cough.

Highlights

  • Chlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest

  • In this study sensitivity and clinical performance seems to be the same for conventional PCR and real-time PCR, but this needs to be confirmed by analysis of more clinical samples

  • We extended the investigation of clinical performance with determination of the PCR efficiency in the samples

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Summary

Introduction

Chlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest. C. pneumoniae infection has been detected by serological methods but PCR is currently viewed as an advantageous alternative since it detects the presence of the DNA of the organism. This allows for an early and clinically relevant diagnosis in contrast to the detection of C. pneumoniae specific antibodies that develop late in the course of the infection. Real-time quantitative PCR in the Lightcycler is a new option among the many PCR assays and it may become a method of choice as it is fast and provides a quantitative measure This is advantageous in a clinical setting because it allows fast diagnosis and fast treatment with relevant antibiotics. The quantitative measure could possibly be used to assess the response to treatment or to assess the state of infection; active infection versus chronic infection

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