Utility of Major Leukocyte Subpopulations for Monitoring Secondary Cytomegalovirus Infections in Renal-Allograft Recipients by PCR

Journal of Clinical Microbiology - Tập 36 Số 4 - Trang 1008-1014 - 1998
Peter Schäfer1, W. Tenschert2, Liana Cremaschi2, K. Gutensohn3, Rainer Laufs1
1Institut für Medizinische Mikrobiologie and Immunologie,1
2Urologische Klinik,2 and
3Abteilung für Transfusionsmedizin,3Universitäts-Krankenhaus Eppendorf, Hamburg, Germany

Tóm tắt

ABSTRACT The feasibility of the major peripheral blood leukocyte (PBL) subsets for use in qualitative and quantitative PCR to monitor secondary cytomegalovirus (CMV) infection and ganciclovir therapy was assessed with 188 blood samples derived from 40 CMV immunoglobulin G-positive renal-allograft recipients. In pp65 antigen-positive patients all leukocyte fractions, but only 79.5% of plasma preparations, were PCR positive. In pp65 antigen-negative samples from patients after antiviral treatment only 7.3% of polymorphonuclear cell (PMNL) samples, but 81.8% of peripheral blood mononuclear cells (PBMC), and 10.9% of plasma samples remained PCR positive. Similarly, in patients with latent infections only 5.0% of PMNL, but 51.7% of PBMC preparations, and 8.0% of plasma samples were PCR positive. Regarding patients with active CMV infection, CMV DNA copy numbers in PMNL correlated significantly with pp65 antigen-positive cell counts before and after onset of ganciclovir therapy. Significant differences in CMV DNA copy numbers in PMNL and plasma were observed (i) between patients with symptomatic infection and those with asymptomatic infection and (ii) between patients with active infection and those with latent infection. In contrast, PBMC harbored equally low CMV DNA levels both in patients with active infection and those with latent infections, and no decline of CMV DNA load in PBMC was observed during antiviral treatment. We conclude that detection of CMV DNA in PMNL, not in PBMC, is associated with active infections and is more sensitive than detection of CMV DNA in plasma. Negative PCR results for PMNL after antiviral therapy indicate recovery, and fewer unwanted positive results occur compared to PBMC and plasma. Therefore, purified PMNL should be preferred for analysis by qualitative CMV PCR to avoid unwanted positive results. The CMV DNA load in PBMC compared with that in PMNL is negligible during active infection, so mixed PBL are sufficient for use in quantitative PCR.

Từ khóa


Tài liệu tham khảo

10.1097/00007890-199307000-00020

10.1093/infdis/167.3.740

10.1128/AAC.41.4.860

10.1002/j.1460-2075.1986.tb04606.x

10.1093/infdis/161.1.31

10.1128/jcm.30.7.1876-1878.1992

10.1128/jcm.32.6.1614-1618.1994

10.1136/jnnp.56.2.211

10.1128/AAC.38.1.38

10.1128/jcm.32.11.2709-2717.1994

10.1128/jcm.30.5.1232-1237.1992

10.1093/infdis/166.6.1236

10.1128/jcm.31.7.1943-1945.1993

10.1099/0022-1317-75-8-1989

10.1093/infdis/167.2.270

10.1099/0022-1317-73-11-2923

10.1007/BF01718831

10.1093/nar/18.22.6733

10.1128/jcm.10.4.533-537.1979

10.1002/jmv.1890440416

Ljungman P. Plotkin S. A. Workshop on CMV disease; definitions, clinical severity scores, and new syndromes.Scand. J. Infect. Dis. Suppl. 99 1995 87 89

10.7326/0003-4819-100-2-222

10.1128/jcm.34.12.2959-2962.1996

10.1128/jcm.32.6.1431-1434.1994

10.1093/infdis/136.5.667

10.1172/JCI113313

10.1172/JCI116059

10.1099/0022-1317-74-12-2699

10.1159/000150496

10.1128/jcm.35.3.741-744.1997

10.1093/infdis/175.2.302

10.1128/jcm.30.9.2359-2365.1992

10.1136/bmj.299.6704.897

10.1099/0022-1317-74-2-265

10.1099/0022-1317-72-9-2059

The T. H. van den Berg A. P. Harmsen M. C. van der Bij W. van Son W. J. The cytomegalovirus antigenemia assay: a plea for standardization.Scand. J. Infect. Dis. Suppl. 99 1995 25 29

10.1016/0882-4010(87)90062-3

10.1128/jcm.34.12.3097-3100.1996

10.1093/infdis/172.2.365

10.1002/jmv.1890430217

10.1128/jcm.33.10.2607-2611.1995

10.1128/jcm.30.2.527-530.1992