Quantitative polymerase chain reaction for human herpesvirus diagnosis and measurement of Epstein–Barr virus burden in posttransplant lymphoproliferative disorder

Human herpesviruses can cause acute diseases such as chicken pox or mononucleosis, but also may reactivate during immunosuppression and result in severe or life-threatening illnesses such as shingles or lymphoproliferative disorders. We report the development and validation of a quantitative PCR met...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 1997-10, Vol.43 (10), p.1843-1849
Main Authors: Bai, Xin, Hosler, Gregory, Rogers, Beverly Barton, Dawson, D. Brian, Scheuermann, Richard H
Format: Article
Language:English
Subjects:
Biological and medical sciences
Hematology
Herpesviridae Infections - diagnosis
Herpesviridae Infections - virology
Herpesvirus 4, Human - isolation & purification
Humans
Investigative techniques, diagnostic techniques (general aspects)
Lymphoproliferative Disorders - virology
Medical sciences
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Polymerase Chain Reaction - methods
Postoperative Complications - diagnosis
Postoperative Complications - virology
Reproducibility of Results
Sensitivity and Specificity
Transplantation
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Summary:Human herpesviruses can cause acute diseases such as chicken pox or mononucleosis, but also may reactivate during immunosuppression and result in severe or life-threatening illnesses such as shingles or lymphoproliferative disorders. We report the development and validation of a quantitative PCR method to measure viral burden for all eight human herpesviruses (HSV1, HSV2, VZV, EBV, CMV, HHV6, HHV7, and KSHV) in patients' samples. The method uses an internal standard that is coamplified with the viral target, allowing quantification of viral genomes in absolute terms (e.g., viral targets/mL of blood) and ruling out false-negative results. We demonstrate that transplant patients with lymphoproliferative disorder carry an EBV viral burden 3 logs higher than nontransplant patients. EBV titers in transplant patients without a lymphoproliferative disorder are between these values. This quantitative PCR method may aid in differentiating clinically significant vs latent viral burden in immunosuppressed patients.
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/43.10.1843