Comparison of Quantitation of Cytomegalovirus DNA by Real-Time PCR in Whole Blood with the Cytomegalovirus Antigenemia Assay

BACKGROUND: Quantitation of cytomegalovirus (CMV) DNA using real-time PCR has been utilized for monitoring CMV infection. However, the CMV antigenemia assay is still the 'gold standard' assay. There are only a few studies in Korea that compared the efficacy of use of real-time PCR for quantitation of CMV DNA in whole blood with the antigenemia assay, and most of these studies have been limited to transplant recipients. METHOD: 479 whole blood samples from 79 patients, falling under different disease groups, were tested by real-time CMV DNA PCR using the Q-CMV real-time complete kit (Nanogen Advanced Diagnostic S.r.L., Italy) and CMV antigenemia assay (CINA Kit, ArgeneBiosoft, France), and the results were compared. Repeatedly tested patients were selected and their charts were reviewed for ganciclovir therapy. RESULTS: The concordance rate of the two assays was 86.4% (Cohen's kappa coefficient value=0.659). Quantitative correlation between the two assays was a moderate (r=0.5504, P<0.0001). Among 20 patients tested repeatedly with the two assays, 13 patients were transplant recipients and treated with ganciclovir. Before treatment, CMV was detected earlier by real-time CMV DNA PCR than the antigenemia assay, with a median difference of 8 days. After treatment, the antigenemia assay achieved negative results earlier than real-time CMV DNA PCR with a median difference of 10.5 days. CONCLUSIONS: Q-CMV real-time complete kit is a useful tool for early detection of CMV infection in whole blood samples in transplant recipients.

Comparison of a rapid cytomegalovirus pp65 antigenemia assay revealed by immunofluorescence to an in-house assay revealed by immunoperoxidase for diagnosis in solid organ transplant recipient patients

Cytomegalovirus (CMV) antigenemia is still one of the two major assays available for diagnosis and monitoring of CMV infections. A commercial rapid test recently available in Brazil for quantification of human cytomegalovirus pp65 antigenemia revealed by immunofluorescence technique was compared with the original in-house method revealed by immunoperoxidase in patients receiving solid organ transplants. Of 80 blood samples tested for CMV antigenemia, 34 (42.5 percent) were positive: commercial assay detected 33 (97 percent) and in-house assay detected 20 (58.8 percent) samples. The numbers of positive cells in the two assays were different, with a median of 4.5 and 12 positive cells obtained by in-house and commercial kit, respectively. Discrepancies between assays occurred in 15 specimens from patients with low-grade antigenemia (median 6 positive cells). The assay-time was reduced in approximately 50 percent compared to in-house methodology. In conclusion, besides comparable results obtained for both assays, the commercial antigenemia assay provides more rapid and sensitive results.

Diagnosis value of PP65 antigenemia assay in febrile children with cytomegalovirus infection / 中国医师进修杂志

Objective To evaluate diagnosis value of PP65 antigenaemia assay(AA) on cytomegalovirus(CMV) infection in febrile children.Method One hundred and twenty-eight febrile children with febrile duration between one to two weeks,their blood preparations and urine aliquots were screened,indirect IF staining(antigen PP65),ELISA(CMV-IgM) and light microscope(urinary cytomegalic inclusion).Three methods above mentioned were used in all patients.Result The positive rate of PP65 antigenaemia,CMV-IgM and urinary cytomegalic inclusion was 17.2%,8.6%,6.3%,respectively.The positive coincidence rate of PP65 antigen with CMV-IgM and PP65 antigen with urinary cytomegalic inclusion was 50%(11/22),36%(8/22),respectively.Nineteen childrens antigenaemia PP65 became negative with patient′s condition improved.Three children′s PP65 antigenaemia remained positive,when they were clinic cured.Two of them became negative after two weeks and one after three weeks.Conclusion Antigenaemia PP65 is effective in early diagnosis of active CMV infection,predicting patient′s condition and providing early intervention and treatment.

Cytomegalovirus Infections after Renal Transplantation / 감염

Cytomegalovirus (CMV) infections commonly develop between the second and sixth month after renal transplantation. The incidence of CMV infection in renal transplant recipients, which depends on their serological status regarding CMV antibody and the use of immunosuppressants, is reported to be 38~67%. However, CMV infection after renal transplantation has been rarely reported in Korea. Recently, we experienced 4 cases of symptomatic CMV infection after renal transplantation, which were diagnosed by CMV antigenemia assay. With the development of more sensitive diagnostic methods, CMV infection may become recognized more commonly among renal transplant recipients.

Congenital Asymptomatic Cytomegalovirus Infection: A Comparison of Specific IgM Antibody Test and pp65 Antigenemia Assay

PURPOSE: It is increasingly important to diagnosis asymptomatic infections which make up a majority (90%) of congenital cytomegalovirus (CMV) infections and that they may have sequeles such as sensorineural hearing loss and mental retardation. Recently antigenemia assay has been developed by using monoclonal antibodies against early structural protein pp65 of CMV. This CMV antigenemia assay seems to be more quicker to diagnosis than conventional viral culture or other tests. In this study, we evaluated the CMV antigenemia assay in neonatal congenital asymptomatic CMV infections comparing it to the CMV specific IgM test that uses enzyme immunoassay. METHODS: From October 1995 to May 1996, 231 normal term newborns delivered with asymptomatic in St. Holy Hospital of Catholic University were included. The CMV antigenemia assay was performed with CMV-vueTM Kit by immunocytochemical staining and the CMV specific IgM test was performed with Enzygnost Anti-CMV/IgM by using an enzyme immunoassay. RESULTS: Three cases (male 2, female 1) were CMV pp65 antigenemia assay positive, but none of them were CMV specific IgM antibody test positive. The CMV pp65 antigenemia assay was more sensitive than CMV specific IgM antibody test for detection of congenital asymptomatic CMV infections by 1.3% and 0%, respectively. CONCLUSION: According to previous results, we suggest that the rate of congenital CMV infections using only CMV specific IgM tests have been underestimated. We recommend the CMV antigenemia assay as the preferred method for more rapid and accurate diagnosis of CMV infections. And congenital asymptomatic CMV infections should be diagnosed and followed up because of possible future sequeles.