An Increase in the Number of Polymorphonuclear Leukocytes Inoculated on Shell-Vial Culture Increases the Sensitivity of This Assay in the Detection of Cytomegalovirus in the Blood of Immunocompromised Patients

Abstract

A prospective study was conducted comparing the sensitivity of the pp65 antigenemia assay (AGA) to that of the shell-vial culture (SVC) inoculated with increasing quantities of polymorphonuclear leukocytes (PMNLs) in the detection of cytomegalovirus (CMV) in peripheral blood. From the cellular suspension, three SVCs were inoculated with 200,000, 400,000, and 800,000 PMNLs, respectively. Of the 201 patients studied, 67 (31.9%) had positive results in one of the two analytic tests (AGA or SVC). In this group, 13 (19.4%) presented a negative AGA assay; 13 (19.4%) an AGA of 1; 13 (19.4%) an AGA of between 2 and 5; and 28 (41.8%) an AGA with a value >6 PMNL-positive × 100,000 PMNLs. The SVC inoculated with 200,000 PMNLs detected the presence of CMV in 42 cases (62.6%); 55 (82%) with 400,000; and 64 (95.5%) with 800,000. Statistically significant differences were observed between the isolation capacities of the SVC inoculated with 200,000 and 400,000, and the SVC inoculated with 800,000 PMNLs ( p = 0.0001). In the comparison of the overall sensitivity of the AGA with that of the SVC with 200,000, the AGA was found to be significantly more sensitive ( p = 0.0052). When comparing with the SVC with 400,000 PMNLs, the two techniques were found to be equally sensitive; and in the comparison with the SVC with 800,000, the culture displayed a greater detection sensitivity ( p = 0.0023). According to these results, it seems evident that the increase in the absolute number of PMNLs inoculated in the SVC leads to a significant increase in the sensitivity of the SVC in the detection of low-level viremia by CMV.