Detection of Epstein-Barr virus DNA by polymerase chain reaction in blood and tissue biopsies from patients with Sjogren's syndrome.
Polymerase chain reaction has been used to detect increased levels of EBV DNA in salivary gland (SG) biopsies and PBL from patients with Sjogren's syndrome (SS). These results suggest that EBV, which has a normal site of latency in a small number of SG epithelial cells, may be reactivated in SS patients and provide a target for immune attack. The great sensitivity of polymerase chain reaction (PCR) and the ability to analyze very small tissue biopsies (37) make this technique well suited for clinical diagnosis. Specific methods to prevent artefactual contamination of tissue biopsy DNA with viral DNA of other samples (i.e., lyophilization of samples before DNA extraction) and the use of an internal positive control (i.e., inclusion of primers for a single copy human gene) during PCR amplification are presented. Since EBV reactivation occurs with markedly increased frequency in patients with lymphoproliferative and immunodeficiency diseases, as well as transplant recipients receiving cyclosporin A (10), rapid methods of viral detection such as PCR may allow better monitoring of medications and early detection of EBV-related lymphomas that may arise in these patients.
Saito
I
I
Department of Basic and Clinical Research, Scripps Clinic, La Jolla, California 92037.
Servenius
B
B
Compton
T
T
Fox
R I
RI
D001483:Base Sequence; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012469:Salivary Glands / Q000032:analysis* / Q000382:microbiology / Q000473:pathology; D012859:Sjogren's Syndrome / Q000378:metabolism* / Q000382:microbiology / Q000473:pathology; D019914:Taq Polymerase
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase