The Assessment of the Association Between Herpesviruses and Subgingival Bacterial Plaque by Real-time PCR Analysis

2018 ◽  
Vol 69 (2) ◽  
pp. 507-510 ◽  
Author(s):  
Sorina Mihaela Solomon ◽  
Ana Maria Filioreanu ◽  
Carmen Gabriela Stelea ◽  
Simona Ionela Grigoras ◽  
Irina Georgeta Sufaru ◽  
...  
Keyword(s):  
Real Time ◽  
Human Cytomegalovirus ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Pathogenic Bacteria ◽  
Periodontal Pocket ◽  
Pcr Analysis ◽  
Barr Virus ◽  
Epstein Barr ◽  
Bacterial Plaque

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.

Detection of Epstein-Barr virus and Human Cytomegalovirus in Colorectal Cancer Tissue by Real Time PCR

2021 ◽  
Vol 24 (05) ◽  
Author(s):  
Bashar A. Abdulhassan ◽  
Asmaa B. Al-obaidi ◽  
Noora M. Kareem ◽  
Haidar A. Shamran
Keyword(s):  
Colorectal Cancer ◽  
Real Time ◽  
Human Cytomegalovirus ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Cancer Tissue ◽  
Colorectal Cancer Tissue ◽  
Barr Virus ◽  
Epstein Barr

scholarly journals Anchor-Based Fluorescent Amplicon Generation Assays (FLAG) for Real-Time Measurement of Human Cytomegalovirus, Epstein–Barr Virus, and Varicella-Zoster Virus Viral Loads

2008 ◽  
Vol 54 (11) ◽  
pp. 1900-1907 ◽  
Author(s):  
Alessandro Di Nicola ◽  
Elisa Ghezzi ◽  
Federico Gillio ◽  
Francesco Zerilli ◽  
Erlet Shehi ◽  
...  
Keyword(s):  
Viral Load ◽  
Real Time ◽  
Human Cytomegalovirus ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Varicella Zoster ◽  
Viral Loads ◽  
Barr Virus ◽  
Epstein Barr ◽  
Diagnostic Sensitivity And Specificity

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.

Characteristics of Epstein-Barr Virus-Associated Hemophagocytic Lymphohistiocytosis (HLH) in European Children.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3883-3883
Author(s):  
Hans-Joachim S. Wagner ◽  
Katrin Beutel ◽  
Marion E. Schneider ◽  
Gritta E. Janka
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Blood ◽  
Hemophagocytic Lymphohistiocytosis ◽  
Epstein Barr Virus ◽  
Blood Analysis ◽  
Pcr Analysis ◽  
Course Of Disease ◽  
Barr Virus ◽  
Epstein Barr

Abstract Hemophagocytic lymphohistiocytosis (HLH) represents a severe sepsis-like disease with massive hypercytokinemia and with proliferating and organ-infiltrating phagocytes leading to cytopenia of different hematopoietic lineages. Besides primary familial cases of HLH, infections can cause secondary forms of HLH, among which Epstein-Barr virus (EBV) infections are the most frequent trigger. Between 2003 and 2005, 10 cases of EBV-associated HLH were analysed at a local subcenter of the international HLH study group. Serial blood samples from these 4 boys and 6 girls (age range: 2–14 years, median of age: 4 years) were examined for EBV load in peripheral blood mononuclear cells (PBMC) and plasma by real-time PCR analysis. EBV loads in both PBMC and plasma were significantly elevated in all HLH patients as compared with healthy controls and also with immunocompetent patients suffering from primary EBV infections. In one patient with excessively elevated EBV load in peripheral blood, analysis of EBV-DNA in separated T and B cells by real-time PCR showed an EBV infection of both cell types. During the course of disease, EBV load in HLH patients reflected the activity of disease. Regarding the clinical course and biochemical markers of disease such as LDH, ferritin or soluble IL2-receptor levels in plasma, patients with EBV-associated HLH did not differ from other patients with HLH. Conclusion: Measurement of EBV-load by real-time PCR facilitates detection of EBV-associated cases of HLH and monitoring of affected patients during the course of disease. EBV load analysis should become part of the initial routine examinations of newly diagnosed HLH patients in order to identify the frequency of EBV-associated forms of HLH. Without molecular diagnosis by means of EBV load measurement in peripheral blood, EBV-associated cases of HLH may be underdiagnosed in non-Asian populations.

scholarly journals Comparison of Commercial Real-Time PCR Assays for Quantification of Epstein-Barr Virus DNA

2005 ◽  
Vol 43 (5) ◽  
pp. 2053-2057 ◽  
Author(s):  
G. Ruiz ◽  
P. Pena ◽  
F. de Ory ◽  
J. E. Echevarria
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Barr Virus ◽  
Pcr Assays ◽  
Epstein Barr

Quantitation of Epstein-Barr virus mRNA using reverse transcription and real-time PCR

2004 ◽  
Vol 74 (4) ◽  
pp. 612-618 ◽  
Author(s):  
Birgit Weinberger ◽  
Annelie Plentz ◽  
Klaus M. Weinberger ◽  
Joachim Hahn ◽  
Ernst Holler ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Barr Virus ◽  
Epstein Barr

scholarly journals Heterogeneous, restricted patterns of Epstein–Barr virus (EBV) latent gene expression in patients with chronic active EBV infection

2001 ◽  
Vol 82 (10) ◽  
pp. 2385-2392 ◽  
Author(s):  
Mikio Yoshioka ◽  
Nobuhisa Ishiguro ◽  
Hiroaki Ishiko ◽  
Xiaoming Ma ◽  
Hideaki Kikuta ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cell ◽  
Real Time ◽  
Real Time Pcr ◽  
Epstein Barr Virus ◽  
Clinical Symptoms ◽  
The Other ◽  
Early Gene ◽  
Barr Virus ◽  
Epstein Barr

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.

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