Analysis of a quantitative PCR assay for CMV infection in liver transplant recipients: an intent to find the optimal cut-off value.

BACKGROUND: Preemptive therapy required highly predictive tests for CMV disease. CMV antigenemia assay (pp65 Ag) has been commonly used for rapid diagnosis of CMV infection. Amplification methods for early detection of CMV DNA are under analysis. OBJECTIVES: To compare two diagnostic methods for CMV infection and disease in this population: quantitative PCR (qPCR) performed in two different samples, plasma and leukocytes (PMNs) and using a commercial diagnostic test (COBAS Amplicor Monitor Test) versus pp65 Ag. STUDY DESIGN: Prospective study conducted in liver transplant recipients from February 2000 to February 2001. RESULTS: Analyses were performed on 164 samples collected weekly during early post-transplant period from 33 patients. Agreements higher than 78% were observed between the three assays. Optimal qPCR cut-off values were calculated using ROC curves for two specific antigenemia values. For antigenemia >or=10 positive cells, the optimal cut-off value for qPCR in plasma was 1330 copies/ml, with a sensitivity (S) of 58% and a specificity (E) of 98% and the optimal cut-off value for qPCR-cells was 713 copies/5x10(6) cells (S:91.7% and E:86%). Using a threshold of antigenemia >or=20 positive cells, the optimal cut-off values were 1330 copies/ml for qPCR-plasma (S 87%; E 98%) and 4755 copies/5x10(6) cells for qPCR-cells (S 87.5%; E 98%). Prediction values for the three assays were calculated in patients with CMV disease (9 pts; 27%). Considering the assays in a qualitative way, the most sensitive was CMV PCR in cells (S: 100%, E: 54%, PPV: 40%; NPV: 100%). Using specific cut-off values for disease detection the sensitivity, specificity, PPV and NPV for antigenemia >or=10 positive cells were: 89%; 83%; 67%; 95%, respectively. For qPCR-cells >or=713 copies/5x10(6) cells: 100%; 54%; 33% and 100% and for plasma-qPCR>or=1330 copies/ml: 78%, 77%, 47%, 89% respectively. CONCLUSIONS: Optimal cut-off for viral load performed in plasma and cells can be obtained for the breakpoint antigenemia value recommended for initiating preemptive therapy with high specificities and sensitivities. Diagnostic assays like CMV pp65 Ag and quantitative PCR for CMV have similar efficiency and could be recommended as methods of choice for diagnosis and monitoring of active CMV infection after transplantation.

Aspergillus antigenemia sandwich-enzyme immunoassay test as a serodiagnostic method for invasive aspergillosis in liver transplant recipients.

BACKGROUND: Invasive aspergillosis (IA) is an important cause of mortality in liver transplant patients. Clinical and microbiological diagnosis is difficult, and it is frequently achieved only after autopsy. Early diagnosis and antifungal therapy could improve the survival of these patients. METHODS: A retrospective case-control study of IA in liver transplant recipients (OLT) was performed to determine the value of the detection of galactomannan Aspergillus antigen in serum using a sandwich-ELISA test (Platelia, Sanofi Diagnostic Pasteur). Stored frozen serum specimens obtained during the posttransplantation period were used. RESULTS: Fourteen cases of IA were diagnosed in 240 OLT recipients (IA incidence: 5.8%) during 5 years with 13 deaths (mortality: 93%). Nine case patients and 33 control patients met the criteria required for being considered "valid" for antigenemia analysis. In five of the nine case patients, a serum sample was positive for Aspergillus antigenemia detection. The median value was 5.7 ng/ml (range: 1.6-6.6). Sensitivity of the test was 55.6%, specificity was 93.9%, the positive predictive value was 71.4%, and the negative predictive value was 88.6%. The likelihood ratio of a positive test was 9.2. CONCLUSIONS: Galactomannan detection in serum could be useful for an early diagnosis of IA in OLT recipients.

What do pharmacists think about New York state's new nonprescription syringe sale program? Results of a survey.

Access to sterile syringes can prevent transmission of blood-borne diseases such as human immunodeficiency virus (HIV) and hepatitis B and C. We conducted survey of attitudes of pharmacists to aid in development of the Expanded Syringe Access Demonstration Program (ESAP) in New York State. ESAP is an HIV prevention initiative that authorizes nonprescription sale of hypodermic needles and syringes by registered pharmacies in New York State beginning January 1, 2001. As part of planning for program implementation, the New York State Department of Health (NYSDOH), in collaboration with the New York State Education Department, conducted mailed survey of all 4, 392 licensed pharmacies in New York State during the summer of 2000. Some surveys (171) were returned as undeliverable. Of the 4,221 eligible respondents, 874 (20.7%) completed surveys were received, of which 574 (65.7%) indicated that their pharmacy would likely participate in ESAP. An additional 11.0% were not sure. Only 139 (15.9%) indicated that they would definitely not participate; 7.4% left this question blank. There were 608 responses to questions on safe disposal practices. Of these, 315 (51.8%) respondents indicated that their pharmacy sold sharps containers, and an additional 29 made them available at no cost. Only 133 (21.9%) respondents to this question did not offer sharps containers and were not interested in doing so. In all, 54 responses indicated that they accepted used hypodermic needles and syringes for disposal. Some (170, 28%) that did not accept sharps for disposal were interested in doing so. More than half (382, 63.0%) did not wish to do so. NYSDOH considered respondent suggestions and minimized ESAP requirements. By March 31, 2001, only 3 months after ESAP became effective, more than half of all licensed pharmacies in New York State were registered for ESAP. Survey results provided useful information to NYSDOH and good indication of likelihood of registration. The high level of pharmacy participation in ESAP may be reflective of NYSDOH attention to issues raised by pharmacists, as well as the direct effects of outreach to pharmacy chains regarding ESAP.