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Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome

BACKGROUND: Limited body of evidence suggests that lipopolysaccharide of C. pneumoniae as well as C. pneumoniae-specific immune complexes can be detected and isolated from human serum. The aim of this study was to investigate the presence of viable elementary bodies of C.pneumoniae in serum samples...

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Autores principales: Petyaev, Ivan M, Zigangirova, Nayilia A, Petyaev, Alexey M, Pashko, Ulia P, Didenko, Lubov V, Morgunova, Elena U, Bashmakov, Yuriy K
Formato: Texto
Lenguaje:English
Publicado: Ivyspring International Publisher 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2894221/
https://www.ncbi.nlm.nih.gov/pubmed/20596362
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author Petyaev, Ivan M
Zigangirova, Nayilia A
Petyaev, Alexey M
Pashko, Ulia P
Didenko, Lubov V
Morgunova, Elena U
Bashmakov, Yuriy K
author_facet Petyaev, Ivan M
Zigangirova, Nayilia A
Petyaev, Alexey M
Pashko, Ulia P
Didenko, Lubov V
Morgunova, Elena U
Bashmakov, Yuriy K
author_sort Petyaev, Ivan M
collection PubMed
description BACKGROUND: Limited body of evidence suggests that lipopolysaccharide of C. pneumoniae as well as C. pneumoniae-specific immune complexes can be detected and isolated from human serum. The aim of this study was to investigate the presence of viable elementary bodies of C.pneumoniae in serum samples of patients with acute coronary syndrome and healthy volunteers. MATERIAL AND METHODS: Serum specimens from 26 healthy volunteers and 56 patients with acute coronary syndrome were examined subsequently by serological (C.pneumoniae-specific IgA and IgG), PCR-based and bacteriological methods. Conventional, nested and TaqMan PCR were used to detect C.pneumoniae genetic markers (ompA and 16S rRNA) in DNA from serum specimens extracted with different methods. An alternative protocol which included culturing high-speed serum sediments in HL cells and further C.pneumoniae growth evaluation with immunofluorescence analysis and TaqMan PCR was established. Pellet fraction of PCR-positive serum specimens was also examined by immunoelectron microscopy. RESULTS: Best efficiency of final PCR product recovery from serum specimens has been shown with specific C. pneumoniae primers using phenol-chloroform DNA extraction protocol. TaqMan PCR analysis revealed that human serum of patients with acute coronary syndrome may contain genetic markers of C. pneumoniae with bacterial load range from 200 to 2000 copies/ml serum. However, reliability and reproducibility of TaqMan PCR were poor for serum specimens with low bacterial copy number (<200 /ml). Combination of bacteriological, immunofluorescence and PCR- based protocols applied for the evaluating HL cells infected with serum sediments revealed that 21.0 % of the patients with acute coronary syndrome have viable forms C.pneumoniae in serum. The detection rate of C.pneumoniae in healthy volunteers was much lower (7.7%). Immunological profile of the patients did not match accurately C.pneumoniae detection rate in serum specimens. Elementary bodies of C.pneumoniae with typical ultrastructural characteristics were also identified in serum sediments using immunoelectron microscopy. Conclusions: Viable forms C. pneumoniae with typical electron microscopic structure can be identified and isolated from serum specimens of the patients with acute coronary syndrome and some healthy volunteers. Increased detection rate of C. pneumoniae in serum among the patients with an acute coronary syndrome may contribute towards enhanced pro-inflammatory status in cardiovascular patients and development of secondary complications of atherosclerosis.
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spelling pubmed-28942212010-07-01 Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome Petyaev, Ivan M Zigangirova, Nayilia A Petyaev, Alexey M Pashko, Ulia P Didenko, Lubov V Morgunova, Elena U Bashmakov, Yuriy K Int J Med Sci Research Paper BACKGROUND: Limited body of evidence suggests that lipopolysaccharide of C. pneumoniae as well as C. pneumoniae-specific immune complexes can be detected and isolated from human serum. The aim of this study was to investigate the presence of viable elementary bodies of C.pneumoniae in serum samples of patients with acute coronary syndrome and healthy volunteers. MATERIAL AND METHODS: Serum specimens from 26 healthy volunteers and 56 patients with acute coronary syndrome were examined subsequently by serological (C.pneumoniae-specific IgA and IgG), PCR-based and bacteriological methods. Conventional, nested and TaqMan PCR were used to detect C.pneumoniae genetic markers (ompA and 16S rRNA) in DNA from serum specimens extracted with different methods. An alternative protocol which included culturing high-speed serum sediments in HL cells and further C.pneumoniae growth evaluation with immunofluorescence analysis and TaqMan PCR was established. Pellet fraction of PCR-positive serum specimens was also examined by immunoelectron microscopy. RESULTS: Best efficiency of final PCR product recovery from serum specimens has been shown with specific C. pneumoniae primers using phenol-chloroform DNA extraction protocol. TaqMan PCR analysis revealed that human serum of patients with acute coronary syndrome may contain genetic markers of C. pneumoniae with bacterial load range from 200 to 2000 copies/ml serum. However, reliability and reproducibility of TaqMan PCR were poor for serum specimens with low bacterial copy number (<200 /ml). Combination of bacteriological, immunofluorescence and PCR- based protocols applied for the evaluating HL cells infected with serum sediments revealed that 21.0 % of the patients with acute coronary syndrome have viable forms C.pneumoniae in serum. The detection rate of C.pneumoniae in healthy volunteers was much lower (7.7%). Immunological profile of the patients did not match accurately C.pneumoniae detection rate in serum specimens. Elementary bodies of C.pneumoniae with typical ultrastructural characteristics were also identified in serum sediments using immunoelectron microscopy. Conclusions: Viable forms C. pneumoniae with typical electron microscopic structure can be identified and isolated from serum specimens of the patients with acute coronary syndrome and some healthy volunteers. Increased detection rate of C. pneumoniae in serum among the patients with an acute coronary syndrome may contribute towards enhanced pro-inflammatory status in cardiovascular patients and development of secondary complications of atherosclerosis. Ivyspring International Publisher 2010-06-10 /pmc/articles/PMC2894221/ /pubmed/20596362 Text en © Ivyspring International Publisher. This is an open-access article distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by-nc-nd/3.0/). Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited.
spellingShingle Research Paper
Petyaev, Ivan M
Zigangirova, Nayilia A
Petyaev, Alexey M
Pashko, Ulia P
Didenko, Lubov V
Morgunova, Elena U
Bashmakov, Yuriy K
Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title_full Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title_fullStr Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title_full_unstemmed Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title_short Isolation of Chlamydia Pneumoniae from Serum Samples of the Patients with Acute Coronary Syndrome
title_sort isolation of chlamydia pneumoniae from serum samples of the patients with acute coronary syndrome
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2894221/
https://www.ncbi.nlm.nih.gov/pubmed/20596362
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