Direct quantification of human cytomegalovirus immediate-early and late mRNA levels in blood of lung transplant recipients by competitive nucleic acid sequence-based amplification

A E Greijer, E A Verschuuren, M C Harmsen, C A Dekkers, H M Adriaanse, T H The, J M Middeldorp

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 10(4) copies per 100 microl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.

Original languageEnglish
Pages (from-to)251-9
Number of pages9
JournalJournal of Clinical Microbiology
Volume39
Issue number1
DOIs
Publication statusPublished - Jan 2001

Cite this

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title = "Direct quantification of human cytomegalovirus immediate-early and late mRNA levels in blood of lung transplant recipients by competitive nucleic acid sequence-based amplification",
abstract = "The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 10(4) copies per 100 microl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.",
keywords = "Antibodies, Viral, Cytomegalovirus, Cytomegalovirus Infections, Humans, Immediate-Early Proteins, Lung Transplantation, Phosphoproteins, RNA, Messenger, RNA, Viral, Self-Sustained Sequence Replication, Viral Matrix Proteins, Viral Proteins, Viremia, Evaluation Studies, Journal Article",
author = "Greijer, {A E} and Verschuuren, {E A} and Harmsen, {M C} and Dekkers, {C A} and Adriaanse, {H M} and The, {T H} and Middeldorp, {J M}",
year = "2001",
month = "1",
doi = "10.1128/JCM.39.1.251-259.2001",
language = "English",
volume = "39",
pages = "251--9",
journal = "Journal of Clinical Microbiology",
issn = "0095-1137",
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Direct quantification of human cytomegalovirus immediate-early and late mRNA levels in blood of lung transplant recipients by competitive nucleic acid sequence-based amplification. / Greijer, A E; Verschuuren, E A; Harmsen, M C; Dekkers, C A; Adriaanse, H M; The, T H; Middeldorp, J M.

In: Journal of Clinical Microbiology, Vol. 39, No. 1, 01.2001, p. 251-9.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Direct quantification of human cytomegalovirus immediate-early and late mRNA levels in blood of lung transplant recipients by competitive nucleic acid sequence-based amplification

AU - Greijer, A E

AU - Verschuuren, E A

AU - Harmsen, M C

AU - Dekkers, C A

AU - Adriaanse, H M

AU - The, T H

AU - Middeldorp, J M

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N2 - The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 10(4) copies per 100 microl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.

AB - The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 10(4) copies per 100 microl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.

KW - Antibodies, Viral

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KW - Cytomegalovirus Infections

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KW - Immediate-Early Proteins

KW - Lung Transplantation

KW - Phosphoproteins

KW - RNA, Messenger

KW - RNA, Viral

KW - Self-Sustained Sequence Replication

KW - Viral Matrix Proteins

KW - Viral Proteins

KW - Viremia

KW - Evaluation Studies

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