Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients

B S N Blank, P L Meenhorst, W Pauw, J W Mulder, W C van Dijk, P H M Smits, F Roeles, J M Middeldorp, J M A Lange

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

OBJECTIVE: To evaluate the value of late-pp67-mRNA nucleic acid sequence-based amplification (NASBA) in comparison to DNA-PCR, blood culture and pp65-antigenemia assay for the detection of human cytomegalovirus (HCMV) disease in HIV-infected patients.

METHODS: The results of pp67-mRNA NASBA, DNA-PCR, culture and pp65-antigenemia assay were compared in 402 whole blood specimens of 98 HIV-infected patients with a low CD4 lymphocyte count who had not yet received highly active antiretroviral therapy (HAART). Thirty-seven samples were obtained from 30 patients with a diagnosis of HCMV disease and 365 samples from 68 patients without HCMV disease.

RESULTS: The highest agreement of test results was observed between pp67-mRNA NASBA and quantitative pp65-antigenemia, with a threshold of nine antigen-positive cells/10(5) leukocytes (kappa-value 0.70, 95% CI=0.58-0.82). The sensitivity of pp67-mRNA NASBA for the diagnosis of HCMV disease (59.3%) was identical to that of the quantitative pp65-antigenemia assay, higher than that of the blood culture (48.2%) but lower than that of the DNA-PCR (77.8%). Pp67-mRNA NASBA (92.3%), quantitative pp65-antigenemia assay (92.3%) and blood culture (93.9%) were highly specific for the diagnosis of HCMV disease and as a result, had a higher positive predictive value (76.2, 76.2 and 76.5%, respectively) than the qualitative DNA-PCR (58.3%) and the qualitative pp65-antigenemia assay (47.6%).

CONCLUSION: pp67-mRNA NASBA, an easy and rapid to perform assay, well-standardised by virtue of co-amplified internal system control RNA, provides a high specificity and positive predictive value for the diagnosis of HCMV disease in HIV-infected patients, comparable to that of the quantitative pp65-antigenemia assay and blood culture.

Original languageEnglish
Pages (from-to)29-38
Number of pages10
JournalJournal of Clinical Virology
Volume25
Issue number1
Publication statusPublished - Jul 2002

Cite this

Blank, B. S. N., Meenhorst, P. L., Pauw, W., Mulder, J. W., van Dijk, W. C., Smits, P. H. M., ... Lange, J. M. A. (2002). Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients. Journal of Clinical Virology, 25(1), 29-38.
Blank, B S N ; Meenhorst, P L ; Pauw, W ; Mulder, J W ; van Dijk, W C ; Smits, P H M ; Roeles, F ; Middeldorp, J M ; Lange, J M A. / Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients. In: Journal of Clinical Virology. 2002 ; Vol. 25, No. 1. pp. 29-38.
@article{72de79a1e236448aad586e9558038eb5,
title = "Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients",
abstract = "OBJECTIVE: To evaluate the value of late-pp67-mRNA nucleic acid sequence-based amplification (NASBA) in comparison to DNA-PCR, blood culture and pp65-antigenemia assay for the detection of human cytomegalovirus (HCMV) disease in HIV-infected patients.METHODS: The results of pp67-mRNA NASBA, DNA-PCR, culture and pp65-antigenemia assay were compared in 402 whole blood specimens of 98 HIV-infected patients with a low CD4 lymphocyte count who had not yet received highly active antiretroviral therapy (HAART). Thirty-seven samples were obtained from 30 patients with a diagnosis of HCMV disease and 365 samples from 68 patients without HCMV disease.RESULTS: The highest agreement of test results was observed between pp67-mRNA NASBA and quantitative pp65-antigenemia, with a threshold of nine antigen-positive cells/10(5) leukocytes (kappa-value 0.70, 95{\%} CI=0.58-0.82). The sensitivity of pp67-mRNA NASBA for the diagnosis of HCMV disease (59.3{\%}) was identical to that of the quantitative pp65-antigenemia assay, higher than that of the blood culture (48.2{\%}) but lower than that of the DNA-PCR (77.8{\%}). Pp67-mRNA NASBA (92.3{\%}), quantitative pp65-antigenemia assay (92.3{\%}) and blood culture (93.9{\%}) were highly specific for the diagnosis of HCMV disease and as a result, had a higher positive predictive value (76.2, 76.2 and 76.5{\%}, respectively) than the qualitative DNA-PCR (58.3{\%}) and the qualitative pp65-antigenemia assay (47.6{\%}).CONCLUSION: pp67-mRNA NASBA, an easy and rapid to perform assay, well-standardised by virtue of co-amplified internal system control RNA, provides a high specificity and positive predictive value for the diagnosis of HCMV disease in HIV-infected patients, comparable to that of the quantitative pp65-antigenemia assay and blood culture.",
keywords = "AIDS-Related Opportunistic Infections, Adult, Antigens, Viral, Antiretroviral Therapy, Highly Active, Cytomegalovirus, Cytomegalovirus Infections, Female, Follow-Up Studies, Gene Amplification, HIV Infections, Humans, Male, Phosphoproteins, Prospective Studies, RNA, Messenger, RNA, Viral, Sensitivity and Specificity, Viral Matrix Proteins, Journal Article",
author = "Blank, {B S N} and Meenhorst, {P L} and W Pauw and Mulder, {J W} and {van Dijk}, {W C} and Smits, {P H M} and F Roeles and Middeldorp, {J M} and Lange, {J M A}",
year = "2002",
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pages = "29--38",
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issn = "1386-6532",
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Blank, BSN, Meenhorst, PL, Pauw, W, Mulder, JW, van Dijk, WC, Smits, PHM, Roeles, F, Middeldorp, JM & Lange, JMA 2002, 'Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients' Journal of Clinical Virology, vol. 25, no. 1, pp. 29-38.

Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients. / Blank, B S N; Meenhorst, P L; Pauw, W; Mulder, J W; van Dijk, W C; Smits, P H M; Roeles, F; Middeldorp, J M; Lange, J M A.

In: Journal of Clinical Virology, Vol. 25, No. 1, 07.2002, p. 29-38.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients

AU - Blank, B S N

AU - Meenhorst, P L

AU - Pauw, W

AU - Mulder, J W

AU - van Dijk, W C

AU - Smits, P H M

AU - Roeles, F

AU - Middeldorp, J M

AU - Lange, J M A

PY - 2002/7

Y1 - 2002/7

N2 - OBJECTIVE: To evaluate the value of late-pp67-mRNA nucleic acid sequence-based amplification (NASBA) in comparison to DNA-PCR, blood culture and pp65-antigenemia assay for the detection of human cytomegalovirus (HCMV) disease in HIV-infected patients.METHODS: The results of pp67-mRNA NASBA, DNA-PCR, culture and pp65-antigenemia assay were compared in 402 whole blood specimens of 98 HIV-infected patients with a low CD4 lymphocyte count who had not yet received highly active antiretroviral therapy (HAART). Thirty-seven samples were obtained from 30 patients with a diagnosis of HCMV disease and 365 samples from 68 patients without HCMV disease.RESULTS: The highest agreement of test results was observed between pp67-mRNA NASBA and quantitative pp65-antigenemia, with a threshold of nine antigen-positive cells/10(5) leukocytes (kappa-value 0.70, 95% CI=0.58-0.82). The sensitivity of pp67-mRNA NASBA for the diagnosis of HCMV disease (59.3%) was identical to that of the quantitative pp65-antigenemia assay, higher than that of the blood culture (48.2%) but lower than that of the DNA-PCR (77.8%). Pp67-mRNA NASBA (92.3%), quantitative pp65-antigenemia assay (92.3%) and blood culture (93.9%) were highly specific for the diagnosis of HCMV disease and as a result, had a higher positive predictive value (76.2, 76.2 and 76.5%, respectively) than the qualitative DNA-PCR (58.3%) and the qualitative pp65-antigenemia assay (47.6%).CONCLUSION: pp67-mRNA NASBA, an easy and rapid to perform assay, well-standardised by virtue of co-amplified internal system control RNA, provides a high specificity and positive predictive value for the diagnosis of HCMV disease in HIV-infected patients, comparable to that of the quantitative pp65-antigenemia assay and blood culture.

AB - OBJECTIVE: To evaluate the value of late-pp67-mRNA nucleic acid sequence-based amplification (NASBA) in comparison to DNA-PCR, blood culture and pp65-antigenemia assay for the detection of human cytomegalovirus (HCMV) disease in HIV-infected patients.METHODS: The results of pp67-mRNA NASBA, DNA-PCR, culture and pp65-antigenemia assay were compared in 402 whole blood specimens of 98 HIV-infected patients with a low CD4 lymphocyte count who had not yet received highly active antiretroviral therapy (HAART). Thirty-seven samples were obtained from 30 patients with a diagnosis of HCMV disease and 365 samples from 68 patients without HCMV disease.RESULTS: The highest agreement of test results was observed between pp67-mRNA NASBA and quantitative pp65-antigenemia, with a threshold of nine antigen-positive cells/10(5) leukocytes (kappa-value 0.70, 95% CI=0.58-0.82). The sensitivity of pp67-mRNA NASBA for the diagnosis of HCMV disease (59.3%) was identical to that of the quantitative pp65-antigenemia assay, higher than that of the blood culture (48.2%) but lower than that of the DNA-PCR (77.8%). Pp67-mRNA NASBA (92.3%), quantitative pp65-antigenemia assay (92.3%) and blood culture (93.9%) were highly specific for the diagnosis of HCMV disease and as a result, had a higher positive predictive value (76.2, 76.2 and 76.5%, respectively) than the qualitative DNA-PCR (58.3%) and the qualitative pp65-antigenemia assay (47.6%).CONCLUSION: pp67-mRNA NASBA, an easy and rapid to perform assay, well-standardised by virtue of co-amplified internal system control RNA, provides a high specificity and positive predictive value for the diagnosis of HCMV disease in HIV-infected patients, comparable to that of the quantitative pp65-antigenemia assay and blood culture.

KW - AIDS-Related Opportunistic Infections

KW - Adult

KW - Antigens, Viral

KW - Antiretroviral Therapy, Highly Active

KW - Cytomegalovirus

KW - Cytomegalovirus Infections

KW - Female

KW - Follow-Up Studies

KW - Gene Amplification

KW - HIV Infections

KW - Humans

KW - Male

KW - Phosphoproteins

KW - Prospective Studies

KW - RNA, Messenger

KW - RNA, Viral

KW - Sensitivity and Specificity

KW - Viral Matrix Proteins

KW - Journal Article

M3 - Article

VL - 25

SP - 29

EP - 38

JO - Journal of Clinical Virology

JF - Journal of Clinical Virology

SN - 1386-6532

IS - 1

ER -