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Original ArticleOpen Access
Rapid Detection of Chlamydia pneumoniae DNA in Peripheral Blood Mononuclear Cells of Coronary Artery Disease Patients by Real-Time Fluorescence PCR
Leowattana W 
,
Mahanonda N ,
Pokum S ,
Poungvarin N
,
Mahanonda N ,
Pokum S ,
Poungvarin N NITHI MAHANONDA, M.D.**,
NARAVAT POUNGVARIN,M.D.*
Several recent reports including serological, pathological and animal studies have asso-
ciated
Chlamydia pneumoniae
with coronary artery disease (CAD). In order to establish whether
chronic
C.
pneumoniae
infection is linked to coronary artery disease, clinical intervention trials
may be needed. However, to detect eligible patients with persistent infection, a reliable diagnostic
marker must be developed for identifying cases and assessing efficacy of antichlamydial therapy.
Moreover, the prevalence of circulating
C.
pneumoniae
DNA in CAD patients varied widely from
previous reports. A real-time PCR has been established by using HL-1 and HR-1 primer to amplify
437 base pairs product. Confirmation of the product was performed on LightCycler by melting
curve analysis of detection probes labeled with LC-Red705. Ninety-five angiographically con-
firmed CAD patients and 104 normal, healthy volunteers were recruited. The mononuclear cell
layer was separated from collected blood and rapid, single step real-time PCR was used to detect
C.
pneumoniae
DNA.
C.
pneumoniae
DNA in peripheral blood mononuclear cells (PBMC) was
found in 17 per cent of 95 CAD patients and 1 per cent of 104 normal healthy volunteers (odds
ratio 20.86, 95% confidence interval 2.71 -160.67, p < 0.0001). There was no association between
C.
pneumoniae
DNA in PBMC and serological status. The rapid, real-time PCR showed a clear-
cut result between positive and negative cases. PBMC-based real-time PCR may be a useful tool
for identifying subjects carrying
C.
pneumoniae
in the circulation or in the vascular wall as well.
It
will be a specific indicator of current infection and will be used as a marker for assessing the
microbiological efficacy of antichlamydial therapy in clinical intervention trials.
Key word :
Chlamydia pneumoniae
DNA, Peripheral Blood Mononuclear Cells and Real-Time
Fluorescence PCR
NARAVAT POUNGVARIN,M.D.*
Several recent reports including serological, pathological and animal studies have asso-
ciated
Chlamydia pneumoniae
with coronary artery disease (CAD). In order to establish whether
chronic
C.
pneumoniae
infection is linked to coronary artery disease, clinical intervention trials
may be needed. However, to detect eligible patients with persistent infection, a reliable diagnostic
marker must be developed for identifying cases and assessing efficacy of antichlamydial therapy.
Moreover, the prevalence of circulating
C.
pneumoniae
DNA in CAD patients varied widely from
previous reports. A real-time PCR has been established by using HL-1 and HR-1 primer to amplify
437 base pairs product. Confirmation of the product was performed on LightCycler by melting
curve analysis of detection probes labeled with LC-Red705. Ninety-five angiographically con-
firmed CAD patients and 104 normal, healthy volunteers were recruited. The mononuclear cell
layer was separated from collected blood and rapid, single step real-time PCR was used to detect
C.
pneumoniae
DNA.
C.
pneumoniae
DNA in peripheral blood mononuclear cells (PBMC) was
found in 17 per cent of 95 CAD patients and 1 per cent of 104 normal healthy volunteers (odds
ratio 20.86, 95% confidence interval 2.71 -160.67, p < 0.0001). There was no association between
C.
pneumoniae
DNA in PBMC and serological status. The rapid, real-time PCR showed a clear-
cut result between positive and negative cases. PBMC-based real-time PCR may be a useful tool
for identifying subjects carrying
C.
pneumoniae
in the circulation or in the vascular wall as well.
It
will be a specific indicator of current infection and will be used as a marker for assessing the
microbiological efficacy of antichlamydial therapy in clinical intervention trials.
Key word :
Chlamydia pneumoniae
DNA, Peripheral Blood Mononuclear Cells and Real-Time
Fluorescence PCR
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