scholarly journals Detection of human cytomegalovirus and Epstein-Barr Virus in symptomatic and asymptomatic apical periodontitis lesions by real-time PCR

Author(s):  
S. Ozbek ◽  
A. Ozbek ◽  
MS. Yavuz
2021 ◽  
Vol 24 (05) ◽  
Author(s):  
Bashar A. Abdulhassan ◽  
Asmaa B. Al-obaidi ◽  
Noora M. Kareem ◽  
Haidar A. Shamran

2008 ◽  
Vol 54 (11) ◽  
pp. 1900-1907 ◽  
Author(s):  
Alessandro Di Nicola ◽  
Elisa Ghezzi ◽  
Federico Gillio ◽  
Francesco Zerilli ◽  
Erlet Shehi ◽  
...  

Abstract Background: Monitoring the human cytomegalovirus (HCMV), Epstein–Barr virus (EBV), or varicella-zoster virus (VZV) viral load is an important factor in the management of immunosuppressed patients, such as recipients of solid-organ or bone marrow transplants. The advent of real-time PCR technologies has prompted the widespread development of quantitative PCR assays for the detection of viral loads and other diagnostic purposes. Methods: The fluorescent amplicon generation (FLAG) technology uses the PspGI restriction enzyme to monitor PCR product generation. We modified the FLAG technology by introducing an accessory oligonucleotide “anchor” that stabilizes the binding of the forward primer to the target sequence (a-FLAG). We developed assays for HCMV, EBV, and VZV that incorporated an internal amplification-control reaction to validate negative results and extensively analyzed the performance of the HCMV a-FLAG assay. Results: The 3 assays performed similarly with respect to reaction efficiency and linear range. Compared with a commercially available kit, the HCMV a-FLAG assay results showed good correlation with calculated concentrations (r = 0.9617), excellent diagnostic sensitivity and specificity (99% and 95%, respectively), and similar values for the linear range (1–107 copies/μL), analytical sensitivity (0.420 copies/μL), and intra- and interassay imprecision. Conclusions: The a-FLAG assay is an alternative real-time PCR technology suitable for detecting and quantifying target-DNA sequences. For clinical applications such as the measurement of viral load, a-FLAG assays provide multiplex capability, internal amplification control, and high diagnostic sensitivity and specificity.


2018 ◽  
Vol 69 (2) ◽  
pp. 507-510 ◽  
Author(s):  
Sorina Mihaela Solomon ◽  
Ana Maria Filioreanu ◽  
Carmen Gabriela Stelea ◽  
Simona Ionela Grigoras ◽  
Irina Georgeta Sufaru ◽  
...  

This study used a real-time PCR analysis to determine possible correlations between periodontal presence of human cytomegalovirus, Epstein-Barr virus and various putative parodontopathogenic bacteria. The study included 18 patients (aged 18-38 years) with aggressive periodontitis, 12 patients (ages 37 to 62) with chronic periodontitis and 30 periodontally healthy subjects (aged 21-54 years). Clinical periodontal evaluation included plaque index, gingival index, percentage of bleeding on probing sites, and probing depth. In each patient, a subgingival bacterial plaque sample was obtained from the deepest periodontal pocket. The real-time fluorogenic PCR detection system was used to determine the number of infectious agents. Human cytomegalovirus was detected in 17 sites with periodontal lesions and in two healthy periodontal sites, and the Epstein-Barr virus was detected in 19 sites with periodontal lesions and in 3 normal periodontal sites. Positive correlations were found between human cytomegalovirus and P. gingivalis, T. forsythia and C. rectus. The Epstein-Barr virus positively correlated with P. gingivalis and T. forsythia. Unfavorable changes in environmental exposure or alteration of the immune system genes can periodically suppress the host�s defence against periodontal aggression, which can then result in reactivation of resident herpesviruses and increased pro-inflammatory mediators followed by an increase in pathogenic bacteria.


2005 ◽  
Vol 43 (5) ◽  
pp. 2053-2057 ◽  
Author(s):  
G. Ruiz ◽  
P. Pena ◽  
F. de Ory ◽  
J. E. Echevarria

2004 ◽  
Vol 74 (4) ◽  
pp. 612-618 ◽  
Author(s):  
Birgit Weinberger ◽  
Annelie Plentz ◽  
Klaus M. Weinberger ◽  
Joachim Hahn ◽  
Ernst Holler ◽  
...  

2001 ◽  
Vol 82 (10) ◽  
pp. 2385-2392 ◽  
Author(s):  
Mikio Yoshioka ◽  
Nobuhisa Ishiguro ◽  
Hiroaki Ishiko ◽  
Xiaoming Ma ◽  
Hideaki Kikuta ◽  
...  

Epstein–Barr virus (EBV) has been shown to infect T lymphocytes and to be associated with a chronic active infection (CAEBV), which has been recognized as a mainly non-neoplastic T-cell lymphoproliferative disorder (T-cell LPD). The systemic distribution of EBV genomes was studied, by real-time PCR, in multiple tissues from six patients with CAEBV, including three patients with T-cell LPD, one patient with B-cell LPD and two patients with undetermined cell-type LPD. There were extremely high loads of EBV genomes in all tissues from the patients. This reflects an abundance of circulating and infiltrating EBV-infected cells and a wide variety of clinical symptoms in the affected tissues. We chose one sample from each patient that was shown by real-time PCR to contain a high load of EBV genomes and examined the expression of EBV latent genes by RT–PCR. EBER1 and EBNA1 transcripts were detected in all samples. Only one sample also expressed EBNA2, LMP1 and LMP2A transcripts in addition to EBER1 and EBNA1 transcripts. Two of the remaining five samples expressed LMP1 and LMP2A transcripts. One sample expressed LMP2A but not LMP1 and EBNA2 transcripts. Another sample expressed EBNA2 but not LMP1 and LMP2A transcripts. The other sample did not express transcripts of any of the other EBNAs or LMPs. None of the samples expressed the viral immediate-early gene BZLF1. These results showed that EBV latent gene expression in CAEBV is heterogeneous and that restricted forms of EBV latency might play a pathogenic role in the development of CAEBV.


1999 ◽  
Vol 37 (1) ◽  
pp. 132-136 ◽  
Author(s):  
Hiroshi Kimura ◽  
Makoto Morita ◽  
Yumi Yabuta ◽  
Kiyotaka Kuzushima ◽  
Koji Kato ◽  
...  

To measure the virus load in patients with symptomatic Epstein-Barr virus (EBV) infections, we used a real-time PCR assay to quantify the amount of EBV DNA in blood. The real-time PCR assay could detect from 2 to over 107 copies of EBV DNA with a wide linear range. We estimated the virus load in peripheral blood mononuclear cells (PBMNC) from patients with symptomatic EBV infections. The mean EBV-DNA copy number in the PBMNC was 103.7 copies/μg of DNA in patients with EBV-related lymphoproliferative disorders, 104.1 copies/μg of DNA in patients with chronic active EBV infections, and 102.2copies/μg of DNA in patients with infectious mononucleosis. These numbers were significantly larger than those in either posttransplant patients or immunocompetent control patients without EBV-related diseases. In a patient with infectious mononucleosis, the virus load decreased as the symptoms resolved. The copy number of EBV DNA in PBMNC from symptomatic EBV infections was correlated with the EBV-positive cell number determined by the in situ hybridization assay (r = 0.842; P < 0.0001). These results indicate that the real-time PCR assay is useful for diagnosing symptomatic EBV infection and for monitoring the virus load.


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