Identification Of Cytomegalovirus Research Articles

COMPARATIVE STUDY BETWEEN VIDAS SYSTEM AND PCR TECHNIQUE IN DIAGNOSIS THE CMV IN ABORTED WOMEN

Cytomegalovirus, also known as CMV, is one of the microorganisms responsible for the highest number of miscarriages and stillbirths among pregnant women around the world. It is also one of the most major causes of fetal mortality. In order to evaluate the effectiveness of the Vidas system and traditional PCR in the identification of cytomegalovirus in pregnant women experiencing complications related to abortion, this research project was designed. This study was successful in collecting sixty serum samples from female participants from the Babylon province, which were then categorized. In the first group there were forty women who were experiencing recurrent miscarriages. In the second group there were twenty women who were healthy and served as a control. According to the findings, forty of the serum samples that were analyzed using the Vidas system were positive, but only thirty of the samples were positive when tested using the PCR approach. The PCR method has a higher sensitivity than the Vidas system when it comes to the detection of cytomegalovirus in pregnant women who have undergone an abortion

Recommendations for Diagnosis and Prevention of Cytomegalovirus-Associated Necrotizing Enterocolitis in Breast-Fed Preterm Infants.

We present the case of a breast-fed preterm infant with postnatally acquired cytomegalovirus (CMV) and severe necrotizing enterocolitis (NEC) associated with CMV. The infant had persistent severe thrombocytopenia with clinical deterioration despite multiple platelet transfusions and maximal medical treatment. Surgical intervention was not feasible owing to the instability of the infant's condition. Upon identification of CMV in urine, intravenous ganciclovir was initiated with significant clinical improvement. We also present a literature review of cases of CMV-related NEC or other gastrointestinal complications in preterm and term infants.

Routine Hematoxylin and Eosin Stain Is Specific for the Diagnosis of Cytomegalovirus Infection in Gastrointestinal Biopsy Specimens.

Gastrointestinal cytomegalovirus (CMV) infection is a serious complication in immunocompromised patients; clinicians often expect expedited results for biopsy specimens. Our goal is to determine the accuracy of identification of CMV on hematoxylin and eosin (H&E) stain. A total of 361 biopsy specimens from 273 patients with suspicion for CMV infection were retrieved. CMV was detected by immunohistochemistry (IHC) in 37 specimens acquired from 33 individual patients (average age = 54 years). Among the CMV-positive patients, 29 (88%) were reported to be immunosuppressed. Colon was the most common affected location. Of 37 CMV-positive specimens by IHC, 28 were positive by H&E (76%), 6 were negative (16%), and 3 were suspicious (8%). Of the 29 positive specimens on H&E, 28 were confirmed by IHC (97%) and 1 was indeterminate (3%). The sensitivity and specificity of H&E were 84% and 94%, respectively; the positive predictive value was 97%, and the negative predictive value was 93% ( P < .00001). Our results show that a preliminary diagnosis of CMV infection, based on H&E stains, can be reported with high specificity and low risk for false-positive results. Suspicious cases should be deferred pending the result of IHC stains.

Identification of cytomegalovirus and human herpesvirus-6 DNA in a patient with corneal endotheliitis

To report the case of a patient with unilateral corneal endotheliitis in which both cytomegalovirus (CMV) and human herpesvirus-6 (HHV6) DNA was identified in the aqueous humor. A 67-year-old man with corneal endotheliitis OD was referred to us for decreased visual acuity. Local corneal stromal edema, pigmented keratic precipitates, a coin-shaped lesion and minimal anterior chamber reaction were observed by slit-lamp biomicroscopy. Cells with owl's eye appearance in the endothelial cell layer were observed by in vivo laser confocal microscopy. The patient had rheumatoid arthritis, which was treated by oral prednisolone and intravenous abatacept. Polymerase chain reaction analysis of aqueous humor samples detected both CMV and HHV6 DNA, but not other HHVs. Treatment with topical ganciclovir and systemic valganciclovir resulted in a clear cornea. A patient with corneal endotheliitis had both CMV and HHV6 DNA identified in the aqueous humor. Although both viruses were identified in this case, clinical manifestations resembled CMV corneal endotheliitis, and it was unclear whether HHV6 could affect the clinical course. Systemic abatacept and corticosteroid therapy might play a positive role in cases with both CMV and HHV6 DNA in this corneal endotheliitis.

Diagnostic yield of the cytomegalovirus (CMV) antigenemia assay and clinical features in solid organ transplant recipients and hematopoietic stem cell transplant recipients with CMV pneumonia

Data are limited on the value of non-invasive diagnostic methods, such as the cytomegalovirus (CMV) antigenemia assay, and the clinical features of CMV pneumonia in patients who have undergone solid organ transplant (SOT) compared with those who have had hematopoietic stem cell transplant (HSCT). All adult patients with suspected CMV pneumonia, who had received SOT or HSCT in a tertiary hospital during a 5-year period, were retrospectively enrolled. CMV pneumonia was defined as clinical and radiographic evidence of pneumonia in association with the isolation of CMV in viral cultures of bronchoalveolar lavage or lung tissue specimens, or with the identification of CMV in lung tissue. In total, 36 patients with CMV pneumonia were identified. Of these, 29 (80%) had received SOT and 7 (20%) had received HSCT. The incidence of CMV pneumonia in the patients with SOT (3.0 per 1000 person-years [95% confidence interval {CI} 0.6-8.7]) was lower than in those with HSCT (17.0 per 1000 person-years [95% CI 9.9-27.2], P=0.003) and CMV-related mortality showed a tendency to have lower mortality in patients with SOT (10% [3/29]) than with HSCT (43% [3/7], P=0.07). The overall sensitivity of the CMV assay (≥1/200,000 leukocytes) in patients with CMV pneumonia was 69% (95% CI 52-84%). The CMV antigenemia test is of limited value in diagnosing CMV pneumonia, given the high cost of false-negative diagnoses of CMV pneumonia.

Identification of cytomegalovirus (CMV)pp65 antigen-specific human monoclonal antibodies using single B cell-based antibody gene cloning from melanoma patients

Recently, because of highly advanced protein engineering technology, beyond the chimeric antibody, highly humanized and fully human antibody development is becoming crucial in the medical field. In the last decade, investigational approaches using clinical samples for fully human antibody production have been performed, but there are still problems with efficiency and accuracy, which should be solved. In the present study, based on novel IgG antibody-measuring ELISA and antibody gene copy number-quantitative PCR, a human single B cell RT-PCR-mediated IgG monoclonal antibody (mAb) gene cloning method was established, and CMVpp65-specific human mAbs were successfully identified. Quantitative PCR for the human IgG mRNA copy number per cell demonstrated that the detection range was 10–250 copies/cell. CMVpp65 +surfaceIgG + B cells were collected from melanoma patients who showed high titers of serum anti-CMVpp65 IgG antibody. RT-PCR was successful in 64% (IGH) and 84% (β-actin) of 88 single B cells. Finally, both IGH and IGL gene amplifications in the same cell were successful in 21 single cells, and 18 IgG antibody genes specific for CMVpp65 antigen were cloned. Four of 13 recombinant human single-chain fragment variable (scFv) antibodies showed strong responses to full-length CMVpp65 protein. These results suggested that the current fully human mAb production procedure through antibody-titer screening by ELISA, single B cell RT-PCR-based antibody gene cloning, and the making of scFv recombinant antibody is an efficient method of therapeutic antibody development.

Cytomegalovirus shedding in the oral cavity of allogeneic haematopoietic stem cell transplant patients

This study was designed to investigate the effect of allogeneic haematopoietic stem cell transplantation (HSCT) on cytomegalovirus (CMV) shedding in the saliva by nested polymerase chain reaction (nested PCR) and its impact on patient survival. One hundred and twenty-four HSCT patients and 124 healthy volunteers were included in the study. Oral swabs were taken before, after 100 days and 1 year of HSCT at the buccal mucosa. Nested PCR was used to detect CMV in the saliva. Time of death after HSCT was displayed, by means of the Kaplan-Meier method, for the following parameters: age and gender of the patient, donor gender, primary disease, stem cell source, platelet number, chronic graft vs host disease (cGVHD) of salivary glands and oral mucosa, and oral CMV shedding. Cox proportional hazards model was used for multivariate survival analysis. While none of the individuals in the control group showed positive swabs for CMV, the frequency of positive CMV oral swabs in patients at day + 100 after HSCT (45.2%) was statistically higher than before (7.2%) and 1 year after HSCT (17.5%). The presence of CMV was not associated with cGVHD and did not have any impact on post-transplant survival. The present study shows that oral CMV shedding occurs after HSCT, especially at day +100 post-transplant. Identification of CMV in the saliva might be important for the early diagnosis of CMV infection in allo-HSTC.

Identification of cytomegalovirus in a liquid‐based gynecologic sample using morphology, immunohistochemistry, and DNA real‐time PCR detection

The viral cytopathic effects of cytomegalovirus (CMV) are rarely encountered in conventional cervical vaginal smears and have never been reported in a liquid-based gynecologic sample (SurePath). We present results of a liquid-based gynecologic sample (SurePath) from an asymptomatic patient with classic CMV-associated granular or dense cyanophilic intracytoplasmic and intranuclear inclusion bodies with a clear surrounding zone. These inclusions were immunohistochemically positive for CMV. The patient also had human papilloma virus (HPV)-associated koilocytes that showed a unique perinuclear immunostaining pattern suggesting coinfection with both CMV and HPV. CMV amplification using real-time polymerase chain reaction (PCR) of DNA extracted from the liquid-based sample confirmed the morphologic and immunohistochemical findings of CMV infection. These observations suggest that a liquid-based preparation can be used to assess CMV infection morphologically, immunohistochemically, and by real-time PCR.

3P-0699 Identification of cytomegalovirus (CMV) in smooth muscle cells from the intima but not the media layer of the aorta obtained from atherosclerotic patients

3P-0699 Identification of cytomegalovirus (CMV) in smooth muscle cells from the intima but not the media layer of the aorta obtained from atherosclerotic patients

Application of fluoroimmunoassay to the identification of low avidity specific IgG against pathogenic human viruses and Toxoplasma gondii

Background: Serological diagnosis of primary viral infections is usually made by detection of specific IgM. In some cases, false positive results (mainly due to crossreactions between closely related viruses) can be obtained. Moreover, some primary infections occur without specific IgM response. Thus, alternative serological approaches are required for diagnosis. Detection of low avidity, specific IgG has been applied as a useful serological marker for diagnosing infections caused by several viruses and Toxoplasma gondii. Objective: The standardization and application of specific IgG avidity assays using a semiautomated solid phase immunoassay (fluoroimmunoassay (FIA)) on the basis of the urea elution principle, for the characterization of low avidity specific IgG against rubella virus, herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalovirus (CMV) and T. gondii. Study design: The method consists of two simultaneous determinations, one as recommended by the manufacturer and the other including a washing step with 8 M urea after the antigen-antibody reaction. A reduction in titer higher than, or equal to, 50% was considered indicative for presence of low avidity specific IgG. Results: When applied to the diagnosis of infections, this method showed sensitivity ranging from 81% to 100%, and absolute specificity. The detection of low avidity specific IgG allowed the differentiation between primary and recurrent infections caused by VZV. Furthermore, it helped in the identification of CMV as the etiological agent of congenital infection in the absence of specific IgM response, as well as in the elucidation of crossreactivity between antigenically related viruses, i.e., VZV and HSV, and Epstein-Barr virus and CMV. Conclusion: FIA can be used for the characterization of the avidity of specific IgG antibody as a diagnostic test in clinical laboratories.

Cytomegalovirus Is Predictably Present in Perineal Ulcers From Immunosuppressed Patients

We examined five consecutive skin biopsy specimens taken from perineal ulcers on immunosuppressed patients. Examination of hematoxylin-eosin-stained sections in conjunction with immunohistochemistry using monoclonal antibodies to early and late viral antigens resulted in identification of cytomegalovirus in all specimens. Cells containing cytomegalovirus were present in the ulcer base and papillary dermis. Herpes simplex virus was identified in three of five specimens. This series demonstrates that cytomegalovirus is predictably present in perineal ulcers from immunocompromised patients, but does not establish this virus as the cause of the ulcers.

Identification of Cytomegalovirus in Bone Marrow Biopsy

The bone marrow trephine biopsy has become a common procedure for evaluating hematopoietic abnormalities. Occasionally, the first clue to an infectious process is obtained from this source. A bone marrow biopsy, done to evaluate pancytopenia in a patient who had received a renal transplant, revealed large intranuclear inclusions diagnostic of cytomegalovirus. Antibody titers were initially negative but rose over the next few weeks to confirm the diagnosis. Even though the bone marrow biopsy may not be the best site for finding CMV inclusions, the careful examination of a random bone marrow biopsy, particularly in an immunosuppressed individual, may contribute to the diagnosis.

Comparison of antiglobulin, direct, and anticomplement immunofluorescent staining for the identification of cytomegalovirus in cultured human fibroblasts

The sequential development of intracellular antigens in cultured human embryonic fibroblasts infected with the AD-169 strain of cytomegalovirus was studied by means of antiglobulin, direct, and anticomplement immunofluorescent staining.Direct immunofluorescent staining, although less sensitive than the antiglobulin or anticomplement methods for the detection of early diffuse nuclear antigens, gave the clearest definition of the typical intracellular inclusions produced by cytomegalovirus.Quantitative complement fixation assays, carried out in parallel with anticomplement staining, showed that complement is fixed strongly by the intracellular antigens which develop in the late stages of infection.