Open AccessIn situ detection of DNA and mRNA of human cytomegalovirus to distinguish different forms of viral infection in leukocytes.
Author(s) -
Barbara Zawilińska,
Katarzyna Bulek,
Jolanta Kopeć,
Magdalena KoszVnenchak
Publication year - 2006
Publication title -
acta biochimica polonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.452
H-Index - 78
eISSN - 1734-154X
pISSN - 0001-527X
DOI - 10.18388/abp.2006_3316
Subject(s) - human cytomegalovirus , biology , in situ hybridization , in situ , virology , virus , cytoplasm , reverse transcriptase , antigen , microbiology and biotechnology , cytomegalovirus , polymerase chain reaction , dna , hybridization probe , messenger rna , nested polymerase chain reaction , gene , herpesviridae , viral disease , immunology , chemistry , genetics , organic chemistry
In situ PCR and in situ reverse transcription PCR (RT-PCR) were applied to discriminate between latent and productive infection of human cytomegalovirus (HCMV) in leukocytes. We investigated 28 samples, in which viral pp65 antigen was detected only in the cytoplasm of leukocytes. Additionally we assayed 12 specimens lacking pp65 antigen. Using nested PCR (nPCR), viral DNA was detected in 27 samples. In six samples the results of nPCR were unreadable due to the presence of polymerase inhibitors. By application of in situ PCR, we were able to confirm the presence of viral DNA in the nucleus and/or cytoplasm. Productive infection was recognized in 20 samples in which transcripts for late viral genes were detected. Among the 20 samples negative by in situ RT-PCR, we recognized phagocytosis of viral particles in eight and the latent form of HCMV infection in five.
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