Comparison of quantitative cytomegalovirus (CMV) PCR in plasma and CMV antigenemia assay: clinical utility of the prototype AMPLICOR CMV MONITOR test in transplant recipients

Abstract

The correlation between the prototype AMPLICOR CMV MONITOR test (Roche Molecular Systems), a quantitative PCR assay, and the cytomegalovirus (CMV) pp65 antigenemia assay was evaluated in transplant recipients. Sequential blood specimens were collected on 29 patients (491 specimens), the leukocyte fraction was tested by CMV antigenemia, and quantitative PCR was performed on plasma specimens. None of the 15 patients (242 specimens) who were antigenemia negative were positive for CMV DNA by PCR, and none of these patients developed active CMV disease. There were 14 antigenemia-positive patients, 8 of whom developed active CMV disease. In all patients, there was a good association between the antigenemia and PCR assays. Ganciclovir-resistant virus was isolated from three patients with active CMV disease. These three patients had persistently elevated levels of antigenemia and CMV DNA by PCR when resistance to ganciclovir developed. This standardized, quantitative CMV PCR assay on plasma has clinical utility for the diagnosis of active disease and in monitoring the response to antiviral therapy in transplant recipients.

FIG. 1

Comparison of CMV antigenemia and quantitative PCR results for the 14 patients who were found to be positive by antigenemia testing. The dashed line (---) represents the negative antigenemia results (no positive cells per 200,000 leukocytes) or a CMV DNA copy number below the limit of detection of the assay (<400 copies/ml). Symbols: ↓, an episode of symptomatic CMV disease; ▵, a specimen that failed to amplify due to the presence of inhibitors; ∗, detection of ganciclovir-resistant virus; ∗∗, detection of foscarnet resistant virus. Patients: A, renal transplant, 25-year-old male, ↓ gastritis; B, renal transplant, 52-year-old male; C, renal transplant, 43-year-old female; D, renal transplant, 62-year-old male; E, bone marrow transplant, 5-year-old male; F, renal transplant, 29-year-old male; G, renal transplant, 49-year-old male, ↓ fever and liver transaminitis; H, renal transplant, 52-year-old female, ↓ (first) fever, fatigue, and leukopenia, ↓ (second) retinitis; I, lung transplant, 40-year-old female, ↓ (first) fever, abdominal pain, liver transaminitis, ↓ (second) retinitis; J, liver transplant, 63-year-old female, ↓ (first) fever and malaise, ↓ (second) leukopenia, low-grade fever and malaise; K, lung transplant, 41-year-old female, ↓ pneumonitis and colitis; L, heart transplant, 56-year-old male; M, liver transplant, 60-year-old male, ↓ fever and fatigue; N, renal transplant, 58-year-old male, ↓ low-grade fever, leukopenia, and liver transaminitis. Bars: □, oral ganciclovir; , intravitreal ganciclovir; ▤, intravenous foscarnet; ▥, oral acyclovir; ■, intravenous ganciclovir; ▧, intravitreal foscarnet; , intravenous acyclovir; , cytogam.

FIG. 1

Comparison of CMV antigenemia and quantitative PCR results for the 14 patients who were found to be positive by antigenemia testing. The dashed line (---) represents the negative antigenemia results (no positive cells per 200,000 leukocytes) or a CMV DNA copy number below the limit of detection of the assay (<400 copies/ml). Symbols: ↓, an episode of symptomatic CMV disease; ▵, a specimen that failed to amplify due to the presence of inhibitors; ∗, detection of ganciclovir-resistant virus; ∗∗, detection of foscarnet resistant virus. Patients: A, renal transplant, 25-year-old male, ↓ gastritis; B, renal transplant, 52-year-old male; C, renal transplant, 43-year-old female; D, renal transplant, 62-year-old male; E, bone marrow transplant, 5-year-old male; F, renal transplant, 29-year-old male; G, renal transplant, 49-year-old male, ↓ fever and liver transaminitis; H, renal transplant, 52-year-old female, ↓ (first) fever, fatigue, and leukopenia, ↓ (second) retinitis; I, lung transplant, 40-year-old female, ↓ (first) fever, abdominal pain, liver transaminitis, ↓ (second) retinitis; J, liver transplant, 63-year-old female, ↓ (first) fever and malaise, ↓ (second) leukopenia, low-grade fever and malaise; K, lung transplant, 41-year-old female, ↓ pneumonitis and colitis; L, heart transplant, 56-year-old male; M, liver transplant, 60-year-old male, ↓ fever and fatigue; N, renal transplant, 58-year-old male, ↓ low-grade fever, leukopenia, and liver transaminitis. Bars: □, oral ganciclovir; , intravitreal ganciclovir; ▤, intravenous foscarnet; ▥, oral acyclovir; ■, intravenous ganciclovir; ▧, intravitreal foscarnet; , intravenous acyclovir; , cytogam.