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

No abstract available.
Colitis/*complications/virology ; Cytomegalovirus Infections/*complications/diagnosis ; *Diabetes Complications ; Humans ; Male ; Middle Aged

Cytomegalovirus (CMV) is an important cause of opportunistic infection in immunocompromised patients. CMV infection occurs as a result of the cell-mediated immunity change in lymphoma patients. Although CMV can cause ulceration anywhere in the gastrointestinal (GI) tract in immunocompromised patients, only a few case reports about CMV GI infection in malignant lymphoma have been documented in literature. Furthermore, it was rare that CMV gastric ulcer with massive bleeding presented as an initial manifestation in a patient who has been not diagnosed lymphoma. We report a case of CMV induced gastric ulcer as an initial manifestation in patient with Hodgkin's disease.
Aged ; Cytomegalovirus ; Cytomegalovirus Infections/*diagnosis/pathology ; Diagnosis, Differential ; Gastroscopy ; Hodgkin Disease/complications/*diagnosis ; Humans ; Male ; Stomach Ulcer/*diagnosis/pathology/virology ; Tomography, X-Ray Computed

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

OBJECTIVETo quantify human cytomegalovirus (HCMV) DNA in the blood and urine of infants of different ages with suspected HCMV infection, and in the breast milk of their mothers, and to evaluate the significance of HCMV DNA detection in the three specimen types in the diagnosis of HCMV infection among infants of different ages.

METHODSA total of 170 infants with suspected HCMV infection were divided into groups A (<28 days; n=43) and B (28 days to 5 months; n=127) according to their ages. Blood and urine were collected from the infants, and breast milk was collected from their mothers. The specimens were examined by fluorescence quantitative PCR for detection of HCMV DNA.

RESULTSIn group A, HCMV DNA detection rates in blood, urine and breast milk were 65.1%, 18.6% and 93.0% respectively. In group B, HCMV DNA detection rates in blood, urine and breast milk were 64.6%, 92.9% and 72.4% respectively. HCMV DNA detection rate in urine in group B was significantly higher than in group A (P<0.01), however, HCMV DNA detection rate in mothers' breast milk in group B was significantly lower than in group A (P<0.01). Among the 82 infants with positive results for blood and urine, the copy number of HCMV DNA in urine was significantly higher than that in blood.

CONCLUSIONSHCMV DNA detection rates in urine and breast milk are different among infants of different ages, so use of suitable specimens according to age is of great significance for improving detection rate.

Animals ; Cytomegalovirus Infections ; diagnosis ; DNA, Viral ; analysis ; blood ; urine ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Milk, Human ; virology ; Pregnancy

BACKGROUNDTo study the composition and significance of the inclusion bodies of human cytomegalovirus (HCMV).

METHODSMicrodissection of inclusion bodies, PCR and Southern blot were adopted to detect DNA, and immunohistochemistry method and catalyzed signal amplification (CSA) were used to detect the different antigens of HCMV.

RESULTSThe inclusion bodies of HCMV were separated from the tissue section of human salivary gland. The fragments amplified by PCR from these dissected inclusion bodies were confirmed to be the DNA of HCMV. With the immunohistochemical method CSA, the immediately early and early antigens of HCMV were detected with monoclonal antibodies DDG9/CCH2, while matrix protein AAC10 was negative in the inclusion bodies.

CONCLUSIONThe ingredient of inclusion bodies of HCMV included the DNA and the antigens expressed in specific stage of the virus.

Antigens, Viral ; analysis ; immunology ; Cytomegalovirus ; genetics ; immunology ; Cytomegalovirus Infections ; diagnosis ; immunology ; virology ; DNA, Viral ; analysis ; genetics ; Humans ; Immunohistochemistry ; Inclusion Bodies ; chemistry ; immunology ; virology ; Microdissection ; Salivary Glands ; chemistry ; immunology ; virology

A 32-year-old man, who had no previous medical history, was hospitalized with 3-week duration of abdominal pain, fever, and watery diarrhea. Initial colonoscopy showed subepithelial hemorrhagic spots throughout the entire colon together with well-circumscribed ulcer around the ileocecal valve. Serologic test disclosed HIV-positive and repeated biopsies at ulcer base finally revealed that the patient had cytomegalovirus ulcer in ileocecal area.
AIDS-Related Opportunistic Infections/complications/*diagnosis ; Adult ; Colitis/complications/diagnosis/*virology ; Colonoscopy ; Cytomegalovirus Infections/complications/*diagnosis ; Humans ; Ileal Diseases/complications/pathology/*virology ; *Ileocecal Valve/pathology ; Male ; Ulcer/complications/pathology/*virology

OBJECTIVETo establish a convenient method for diagnosis of active human cytomegalovirus (HCMV) infection in renal transplant recipients.

METHODSThe expression of HMCV phosphoprotein 65 (HCMV pp65) antigen in the peripheral blood leucocytes was detected by catalyzed signal amplification (CSA), and the results were compared with those by detection of the serum HCMV pp65 using anti-HCMV-IgM.

RESULTSOf the 100 renal transplant recipients, 47 were positive for HCMV pp65 in the peripheral blood leucocytes, with the antigen-positive cell number of (71-/+45)/2x10(5) WBC, and 41 recipients were positive for anti-HCMV-IgM. In the 29 recipients presenting symptoms of HCMV infection, the antigen-positive cell number averaged (83-/+46)/2x 10(5) WBC. All the 50 healthy control subjects showed negative results of pp65 and anti-HCMV-IgM. The diagnostic sensitivity, specificity, positive predictive value and negative predictive value of this leukocyte-based HCMV pp65 detection were 100%, 74.6%, 61.7% and 100%, respectively.

CONCLUSIONThis method provides a sensitive and convenient approach for diagnosis of HCMV infection, which may also provide evidence for adequate administration of the antiviral treatment.

Adult ; Antigens, Viral ; analysis ; Cytomegalovirus ; Cytomegalovirus Infections ; diagnosis ; virology ; Female ; Humans ; Kidney Transplantation ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; Nucleic Acid Amplification Techniques ; methods ; Phosphoproteins ; analysis ; Predictive Value of Tests ; Sensitivity and Specificity ; Transplantation, Homologous ; Viral Matrix Proteins ; analysis

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.
Adolescent ; Adult ; Aged ; Conserved Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; diagnosis ; virology ; DNA, Viral ; blood ; Female ; Humans ; Immunosuppressive Agents ; blood ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Polyomavirus ; genetics ; isolation & purification ; Polyomavirus Infections ; diagnosis ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Species Specificity ; Tacrolimus ; blood ; Time Factors ; Tumor Virus Infections ; diagnosis ; virology ; Viral Load ; Young Adult