OBJECTIVETo investigate the clinical and pathologic features of gastrointestinal cytomegalovirus (CMV) disease, and the histological detection of CMV.

METHODSForty-one gastrointestinal tissues were obtained from 35 patients suspected for gastrointestinal CMV disease. The pathologic changes of these specimens were evaluated by immunohistochemistry and in situ hybridization for CMV, and these were compared to serologic detection methods.

RESULTSCMV was detected in 23/41 tissues. There were 32 serologic CMV DNA tests at the time of biopsies or operations. Compared with histological detection, the sensitivity and specificity of serologic CMV DNA tests were 73.7% and 69.2% respectively.

CONCLUSIONSGastrointestinal histological detection of CMV has important clinical application. HE staining combined with immunohistochemistry and/or in situ hybridization detection is a reliable method to detect CMV.

Biopsy ; Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; diagnosis ; Gastrointestinal Diseases ; virology ; Humans ; Immunohistochemistry ; In Situ Hybridization

OBJECTIVETo evaluate the subgingival prevalence of human cytomegalovirus (HCMV), Epstein-Barr virus-1 (EBV-1) in chronic periodontitis (CP) patients before and after treatment and to analyze the relationship between the prevalent variance and periodontal clinical parameters.

METHODSGingival crevicular fluids of 13 CP patients were collected at baseline, 2 weeks, 2 months and 4 months after periodontal mechanical treatment. HCMV and EBV-1 were detected using nested polymerase chain reaction (n-PCR).

RESULTSThe plaque index (PLI), probing depth (PD) and bleeding index (BI) of CP patients at 2 months, 4 months after periodontal mechanical treatment were evidently lower than before treatment, P < 0.01. These parameters at 4 months after treatment were higher than at 2 months, the differences were significant, P < 0.05. The prevalence of HCMV and EBV in CP patients was 42% (33/78), 14% (11/78). EBV and HCMV were mostly coexistent in the same site [9 sites HCMV(+) in 11 EBV positive sites]. The sites of HCMV(+) and EBV(+) were almost deep pockets. Thirteen of 14 sites with deep pockets were HCMV(+), 9 sites were deep pockets in 11 sites EBV(+). The prevalence of HCMV and EBV (8% and 0 respectively) at 2 weeks was the lowest in all four time points. The prevalence of HCMV and EBV at 2 weeks, 2 months and 4 months following treatment was significantly lower than baseline (P < 0.01), but the prevalence of HCMV (15%) at 2 months after treatment was higher than at 2 weeks (8%), the difference was not significant (P = 0.133).

CONCLUSIONSHerpesviruses may play a role in the development of CP. The changes of the prevalence of herpesviruses before the changes of clinical parameters could be detected after periodontal mechanical treatment. The patients should be re-evaluated and re-treated within 2 months after treatment.

Chronic Periodontitis ; therapy ; Cytomegalovirus ; isolation & purification ; Gingival Crevicular Fluid ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans

OBJECTIVETo investigate human cytomegalovirus (HCMV) and Epstein-Barr virus-type 1 (EBV-1) in GCF and saliva during experimental gingivitis in Chinese young subjects and to evaluate the effect of the virus in the initial stage of gingival inflammation.

METHODSGCF of 14 and 45 and saliva without stimulating in 11 Chinese young males with healthy gingiva were collected at baseline (day 0), day 7, 14 and 21 after stopping oral hygiene and day7 after reestablishing oral hygiene (day 28). DNA of HCMV and EBV-1 were detected by nested-polymerase chain reaction (n-PCR) at the times mentioned above.

RESULTSHCMV was detected in GCF of 4 subjects at baseline, 4 subjects at day 7, 3 subjects at day 14 and 2 subjects at day 21 while the subjects were different. At day 28 HCMV could not be detected. EBV-1 was not detectable in GCF during the experimental gingivitis. HCMV was detected in saliva in 4 subjects and EBV-1 was in 3 subjects. And there is no relationship between the detection of the herpesviruses and the clinical parameters as well.

CONCLUSIONWe suggest that HCMV and EBV-1 are not the important factors during the initial stage of gingival inflammation.

Adult ; Cytomegalovirus ; isolation & purification ; Gingival Crevicular Fluid ; virology ; Gingivitis ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Young Adult

Standardized cytomegalovirus (CMV) DNA quantification is important for managing CMV disease. We evaluated the performance of the Real-Q CMV Quantification Kit (Real-Q assay; BioSewoom, Korea) using whole blood (WB), with nucleic acid extraction using MagNA Pure 96 (Roche Diagnostics, Germany). Real-time PCR was performed on two platforms: the 7500 Fast real-time PCR (7500 Fast; Applied Biosystems, USA) and CFX96 real-time PCR detection (CFX96; Bio-Rad, USA) systems. The WHO international standard, diluted with CMV-negative WB, was used to validate the analytical performance. We used 90 WB clinical samples for comparison with the artus CMV RG PCR kit (artus assay; Qiagen, Germany). Limits of detections (LODs) in 7500 Fast and CFX96 were 367 and 479 IU/mL, respectively. The assay was linear from the LOD to 10(6) IU/mL (R2 ≥0.9886). The conversion factors from copies to IU in 7500 Fast and CFX96 were 0.95 and 1.06, respectively. Compared with the artus assay, for values <1,000 copies/mL, 100% of the samples had a variation <0.7 log10 copies/mL; >1,000 copies/mL, 73.3% and 80.6% of samples in 7500 Fast and CFX96, respectively, had <0.5 log10 copies/mL. The Real-Q assay is useful for quantifying CMV in WB with the two real-time PCR platforms.
Cytomegalovirus/*genetics/isolation & purification ; Cytomegalovirus Infections/diagnosis/virology ; DNA, Viral/*blood/metabolism ; Humans ; Limit of Detection ; Reagent Kits, Diagnostic ; *Real-Time Polymerase Chain Reaction

Human cytomegalovirus (CMV) infection may cause life-threatening complications and lead to failure in transplantation. So, it is very important to explore the laboratory methods which can detect the CMV sufficiently early and accurately. This review discusses methodological aspects of quantitative CMV assays with emphasis on recently developed antigen detection assays and molecular biological methods.
Antigens, Viral ; blood ; Bone Marrow Transplantation ; adverse effects ; methods ; Cytomegalovirus ; immunology ; isolation & purification ; Cytomegalovirus Infections ; diagnosis ; etiology ; virology ; Humans ; Transplantation, Homologous

Cytomegalovirus infection is extremely common in the population, especially for newborns. Congenital CMV infection may cause central nervous system damage and other related diseases, thus potentially harmful. At home and abroad, some related research had been carried out on the incidence of disease, and a variety of detection methods had been developed. In this paper, the current situation of congenital cytomegalovirus infection and detection method is reviewed.
Antibodies, Viral ; blood ; Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; congenital ; diagnosis ; DNA, Viral ; blood ; Female ; Humans ; Infant, Newborn ; Polymerase Chain Reaction ; Pregnancy

OBJECTIVETo investigate human cytomegalovirus (HCMV) glycoprotein B (gB) genotypes and clinical features in neonates with congenital infections.

METHODSUrine samples were obtained from 67 neonates with HCMV infection confirmed by polymerase chain reaction (PCR). The gB gene fragment was amplified by nested PCR. HCMV gB genotyping was detected by restriction fragment length polymorphism.

RESULTSIn all these cases, the most prevalent genotype was gBl (50.7%), followed by gB3 (23.9%), gB2 (17.9%), and gBl/gB3 coinfection (7.5%); gB4 was not found. Moreover, gB1 was more prevalent in infants with liver damage (27/37, 73.0%) than in other symptomatic infants without liver damage (13/30, 43.3%; P < 0.05).

CONCLUSIONThe gBI genotype is the most prevalent in infants with congenital symptomatic HCMV disease, especially in those with liver damage, followed by genotypes gB3, gB2, and gB4.

Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; urine ; virology ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Infant, Newborn, Diseases ; virology ; Male ; Viral Envelope Proteins ; genetics ; urine

OBJECTIVE: To investigate the relationship between MICA*008/A5.1 allele and human cytomegalovirus (HCMV) infection in kidney transplanted donees of Hunan Han nationality. METHODS: The MICA*008/A5.1 allele based on 91 kidney transplanted donees and 81 unrelated normal individuals of Han nationality in Hunan Province were analyzed by PCR/SSP assay. At the same time, anti-HCMV antibody IgM was detected in the serum by ELISA method. RESULTS: The positive rate of MICA*008/A5.1 allele was significantly higher in the control group (56.79%) than that in the kidney transplanted donee group (34.07%) (P <0.05). The infection rate of HCMV in those individuals whose genotype was MICA*008/A5.1 (-) was significantly higher than that in the MICA*008/A5.1(+). CONCLUSION The individual whose genotype is MICA*008/A5.1 (+) is not liable to HCMV infection, but the individual whose genotype is MICA*008/A5.1 (-) is liable to HCMV infection.
Alleles ; Antibodies, Viral ; blood ; China ; Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; genetics ; Female ; Genotype ; Histocompatibility Antigens Class I ; genetics ; Humans ; Immunoglobulin M ; blood ; Kidney Transplantation ; adverse effects ; Male

BACKGROUND: Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are increasingly important in immunocompromised patients. Nucleic acid extraction methods could affect the results of viral nucleic acid amplification tests. We compared two automated nucleic acid extraction systems for detecting CMV and EBV using real-time PCR assays. METHODS: One hundred and fifty-three whole blood (WB) samples were tested for CMV detection, and 117 WB samples were tested for EBV detection. Viral nucleic acid was extracted in parallel by using QIAsymphony RGQ and QIAcube (Qiagen GmbH, Germany), and real-time PCR assays for CMV and EBV were performed with a Rotor-Gene Q real-time PCR cycler (Qiagen). Detection rates for CMV and EBV were compared, and agreements between the two systems were analyzed. RESULTS: The detection rate of CMV and EBV differed significantly between the QIAsymphony RGQ and QIAcube systems (CMV, 59.5% [91/153] vs 43.8% [67/153], P=0.0005; EBV, 59.0% [69/117] vs 42.7% [50/117], P=0.0008). The two systems showed moderate agreement for CMV and EBV detection (kappa=0.43 and 0.52, respectively). QIAsymphony RGQ showed a negligible correlation with QIAcube for quantitative EBV detection. QIAcube exhibited EBV PCR inhibition in 23.9% (28/117) of samples. CONCLUSIONS: Automated nucleic acid extraction systems have different performances and significantly affect the detection of viral pathogens. The QIAsymphony RGQ system appears to be superior to the QIAcube system for detecting CMV and EBV. A suitable sample preparation system should be considered for optimized nucleic acid amplification in clinical laboratories.
Automation ; Cytomegalovirus/*genetics/isolation & purification ; Cytomegalovirus Infections/diagnosis/*virology ; DNA, Viral/*blood/isolation & purification/metabolism ; Herpesvirus 4, Human/*genetics/isolation & purification ; Humans ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the associations between Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) infections and the methylation levels of PTEN and hTERT genes and explore their roles in children with acute lymphoblastic leukemia (ALL).

METHODSBlood samples from 100 children with newly diagnosed acute lymphoblastic leukemia were centrifuged for serological detection of EBV and HCMV, and the patients were divided accordingly into EBV-infected group (n=20), HCMV-infected group (n=14), EBV and HCMV co-infected group (n=41), and non-infected group (control group, n=15). DNA was extracted from peripheral blood mononuclear cells (PBMCs) and modified with bisulfite ammonia sodium. The methylation levels of the promoters of PTEN and hTERT genes were detected with methylation-specific polymerase chain reaction (MS-PCR).

RESULTSCompared with those in non-infected group and EBV- or HCMV-infected group, the methylation levels of PTEN gene in the co-infected group were significantly decreased (P<0.05) while the methylation levels of hTERT gene significantly increased (P<0.05).

CONCLUSIONIn children with acute lymphoblastic leukemia, EBV and HCMV co-infection cause changes in the methylation levels of PTEN and hTERT. These results may be associated with epigenetic changes caused by viral infections, and further studies are needed to further verify this hypothesis.

Child ; Coinfection ; Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; DNA Methylation ; Epstein-Barr Virus Infections ; Herpesvirus 4, Human ; isolation & purification ; Humans ; PTEN Phosphohydrolase ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; genetics ; virology ; Promoter Regions, Genetic ; Telomerase ; genetics ; metabolism