Comparison of Cell Culture with Three Conventional Polymerase Chain Reactions for Detecting Chlamydophila pneumoniae in
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Comparison of Cell Culture with Three Conventional Polymerase Chain Reactions for Detecting Chlamydophila pneumoniae in Adult’s Pharyngotonsillitis Aldo Stivala1 · Carlo Genovese1,2 · Claudia Bonaccorso1 · Valentina Di Salvatore1 · Giulio Petronio Petronio3 · Adriana Garozzo1 · Mario Salmeri1 Received: 27 November 2019 / Accepted: 23 June 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Chlamydophila pneumoniae is an intracellular pathogen responsible for respiratory tract infections. The isolation of the microorganism from clinical specimens is essential for a diagnosis. However, the identification of C. pneumoniae by cell cultures is very difficult besides strongly depending on the sample conditions. The study aimed to investigate, in adult patients with pharyngotonsillitis, the frequency of Chlamydophila pneumoniae detection by cell cultures and three conventional PCRs (a conventional PCR targeting the 16S rRNA gene and two nested PCRs, targeting the 16S rRNA gene and the ompA gene, respectively). The presence of chlamydial inclusion in cell cultures was observed in 11/94 samples (11.70%) by IFA. C. pneumoniae DNA was detected in 12/94 (12.76%) specimens by the 16S rRNA gene nested PCR, 4/94 (4.26%) by ompA gene nested PCR, and in 2/94 (2.13%) by 16S rRNA single-step PCR. Our data show poor agreement between the three applied DNA-amplification methods; in fact, only 16S rRNA gene nested PCR showed a statistically significant difference. Moreover, this result allowed us to achieve a definitive confirmation of the previous finding and to avoid the risk of an overestimation of the C. pneumoniae as a pathogen in pharyngotonsillitis.
Introduction Chlamydophila pneumoniae (C. pneumoniae) is an obligate intracellular bacterium, due to its inability to synthesize ATP. It has a life cycle with two developmental forms, elementary body (EB) and reticulate body (RB), that represent the infective and the reproductive form, respectively. C. pneumoniae is mainly recognized as a respiratory pathogen but also associated with other chronic diseases such as Aldo Stivala and Carlo Genovese have contributed equally to this work. * Carlo Genovese [email protected] 1
Department of Biomedical and Biotechnological Sciences, University of Catania, Via Santa Sofia 97, 95123 Catania, Italy
2
Nacture S.R.L, Spin-Off University of Catania, Via Santa Sofia 97, 95123 Catania, Italy
3
Department of Medicine and Health Sciences “Vincenzo Tiberio”, University of Molise, Via Francesco de Sanctis 1, 86100 Campobasso, Italy
atherosclerosis [1, 34], multiple sclerosis [26], and Alzheimer’s disease [11]. C. pneumoniae infections are widespread in human populations. Seroepidemiological data show that 50–70% of the adult population is exposed to this pathogen during their lifetime [25, 37]. C. pneumoniae is considered an unconventional pathogen in pharyngotonsillitis. Indeed, it is well known that the most common, non-viral, etiological agent of pharyngotonsillitis is Streptococcus pyogenes. Neverth
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