Correlation of cytomegalovirus (CMV) detection in urine by tissue culture virus isolation, early antigen immunofluorescence test and nucleic acid hybridization

Meyer, G; Enders, Gisela

doi:10.1007/bf01644090

Cited by 5 publications

(7 citation statements)

References 29 publications

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“…Isolation of virus from urine in tissue culture during the first week of life the standard procedure for detecting congenital CMV infection. However, with a minimal incubation time of 4 weeks and a mean isolation time of 10-16 days, the technique is too slow (3). A rapid and reliable test to detect congenital CMV infection is essential.…”

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confidence: 99%

Detection of congenital cytomegalovirus infection in Chinese newborn infants using polymerase chain reaction

Tsai

Shih

et al. 1996

Acta Paediatrica

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Detection ofcongenital cytomegalovirus infection in Chinese newborn infants using polymerase chain reaction. Acta Prediatr 1996;85: 1241-3. Stockholm. ISSN 0803-5253 Polymerase chain reaction (PCR) amplification was used to detect cytomegalovirus (CMV) in 1000 urine specimens from Chinese newborns for defining the incidence of congenital CMV infection in the Chinese population. The major immediate-early and the late antigen genes of CMV were amplified and detected by gel electrophoresis. There were 18 congenitally infected infants found when tests were performed with one or both primer pairs. Comparing with tissue culture, PCR of both primer sets provided a sensitivity of 94%. a specificity of 100% and a predictive value of positive result of 100%.

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mentioning

confidence: 99%

Detection of congenital cytomegalovirus infection in Chinese newborn infants using polymerase chain reaction

Tsai

Shih

et al. 1996

Acta Paediatrica

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“…For standardisation of the PCR method it is essential to compare results with those of conventional techniques such as virus isolation. Sensitivity of PCR using primers from the IE1 region (Exon 3/4) was not significantly greater than that of hybridisation [5]. The low sensitivity may be explained by the large size of the amplicons (530 base pairs) or the presence of inhibitors of Taqpolymerase in urine specimens.…”

Section: Discussionmentioning

confidence: 87%

“…Cells were examined under an inverted microscope, cells positive for HCMV immediate early antigen exhibited intensely stained brownish nuclei. Virus isolation in cell culture: Cell culture was performed according to standard tissue culture techniques [5]. After appearance of a specific cytopathic effect (CPE), HCMV infection was confirmed with mAB against an HCMV late protein (NEA-9220, DuPont).…”

Section: Cells and Virus"mentioning

confidence: 99%

“…With the advent of effective antiviral therapy -also in cases of severe congenital infection in newborns -fast and reliable assays for HCMV detection are required. Virus detection in fibroblast cultures by HCMV-specific cytopathic effect (CPE) is still considered to be "gold standard", but with amean isolation time of 10-16 days this technique is too sl0w [5,6]. More rapid assays have been described for detection of early antigens with monoclonal antibodies in fibroblasts and for detection of viral DNA by polymerase chainlreaction (PCR) [7,8,9].…”

Section: Introductionmentioning

confidence: 99%

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Detection of human cytomegalovirus in urine samples by cell culture, early antigen assay and polymerase chain reaction

Daiminger¹,

Schalasta²,

Betzl³

et al. 1994

Infection

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With the advent of effective therapy rapid, sensitive and reliable assays for diagnosis of human cytomegalovirus (HCMV) infections are required. In a total of 1,928 urine samples, detection of HCMV-immediate early antigen in a spin amplified microplate culture by a monoclonal antibody and immunoperoxidase staining (EA-assay) was compared with virus isolation in cell culture. Sensitivity of the EA assay was 85.5% and specificity was 99.5% compared with virus isolation. Overall agreement of both assays was 97.8%. In addition, in 235/1,928 urine samples amplification of HCMV-DNA was performed by means of polymerase chain reaction (PCR) using primers from the immediate early (IE1) gene region and 141/1,928 using primers from the late region (LA). The sensitivity of PCR compared with virus isolation was 67.8% for IE1 primers and 94.1% for LA primers (statistical significance: p < 0.01, Chi-square-test). Overall agreement between virus isolation and PCR was 88.5% for IE1-PCR and 84.4% for LA-PCR. Discordant results were more often found in adults with acute infection and immunocompromised patients than in infants.

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“…Passage für Flachbodenröhrchen-und Mikroplattenkulturen verwendet. Die Anzucht und Zellhaltung erfolgte mit den dafür üblichen Nährmedien [9].…”

Section: Materials Und Methodenunclassified

Vermeidung der zytotoxischen Wirkung von Urinproben im CMV-Early-Antigen-Test durch Ultrafiltration

Daiminger¹,

Enders²

1993

LaboratoriumsMedizin

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Zusammenfassung: Schnelle und zuverlässige Methoden zum Erregernachweis für die Diagnose der Zytomegalie (CMV)-lnfektion sind insbesondere bei der Frage nach antiviraler Therapie von Bedeutung. Der CMV-Early-Antigen-Test (Bestimmung von CMV Frühantigen in Zentrifugationskultur mittels monoklonaler Antikörper) erlaubt den Nachweis von infektiösen Virus schon nach 36 Stunden. Es zeigte sich jedoch, daß etwa 14% aller untersuchten Urine zytotoxisch auf die Zentrifugationskultur wirkten und somit nicht auswertbar waren. Deshalb wurden 160 ausgewählte Urinproben, darunter 73 toxische und 23 leicht toxisch wirkende Proben, mit und ohne vorausgegangene Ultrafiltration in den CMV-Early-Antigen-Test eingesetzt. Es konnte gezeigt werden, daß diese Methode, bei der niedermolekulare, eventuell toxisch wirkende Substanzen entfernt werden, geeignet ist, die zytotoxische Wirkung der Urinproben komplett aufzuheben. Zur Kontrolle der Reproduzierbarkeit wurden die Ergebnisse von 87 auswertbaren Urinen mit und ohne Ultrafiltration verglichen, die Übereinstimmung beider Methoden lag bei 98,8%. Die eingesetzte Ultrafiltrationstechnik ist eine wirksame Methode, um die Zytotoxizität von Urinproben aufzuheben und stellt somit eine weitere Optimierung der CMV-Schnelldiagnostik dar. Schlüsselwörter: CMV-Early-Antigen-Test -Zelltoxizität von Urinproben -Ultra filtration Summary: With the advent ofantiviral therapy, rapid and reliable assays forcytomegalovirus (CMV) detection are required.In fibroblast cultures, detection of CMV early antigen with monoclonal antibody after Iow speed centrifugation (CMV early antigen assay) can be performed after 36 hours. However, in routine diagnosis, evaluation was not possible in 14% of all urine samples due to cell toxicity in the early antigen assay. Therefore, 160 preselected urine samples, containing 73 cytotoxic and 23 slightly toxic probes, were also tested with and without prior Ultrafiltration.We have shown that this method, which removes Iow molecular weight, potentially toxic substances, is able to overcome these toxic effects. In all instances, an intact cell monolayer was maintained after Ultrafiltration and evaluation of the test was possible. To determine reproducibility, 87 non toxic urine samples were tested before and after Ultrafiltration. Concordance of both methods was 98,8%. 566 Lab.med. 17: 566 (1993) Brought to you by | University of Arizona Authenticated Download Date | 5/31/15 4:51 PM OriginalieUltrafilträtion is an effective method of preventing cell toxicity of urine samples in the CMV early antigen assay and therefore enables further optimization of rapid diagnosis of CMV infection.

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Correlation of cytomegalovirus (CMV) detection in urine by tissue culture virus isolation, early antigen immunofluorescence test and nucleic acid hybridization

Cited by 5 publications

References 29 publications

Detection of congenital cytomegalovirus infection in Chinese newborn infants using polymerase chain reaction

Detection of congenital cytomegalovirus infection in Chinese newborn infants using polymerase chain reaction

Detection of human cytomegalovirus in urine samples by cell culture, early antigen assay and polymerase chain reaction

Vermeidung der zytotoxischen Wirkung von Urinproben im CMV-Early-Antigen-Test durch Ultrafiltration

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