A rapid, sensitive, and specific assay to detect mumps virus RNA directly from clinical specimens using a real-time PCR assay was developed. The assay was capable of detecting five copies of standard plasmid containing cDNA from the mumps virus F gene. No cross-reactions were observed with other members of Paramyxoviridae, or with viruses or bacteria known to be meningitis pathogens. Seventy-three clinical samples consisting of throat swabs collected from patients with parotitis, and cerebrospinal fluid (CSF) collected from patients with aseptic meningitis, were examined with a real-time PCR assay developed by the authors, reverse-transcription nested-PCR (RT-n-PCR), and virus isolation using cell culture. Like the RT-n-PCR assay, the real-time PCR assay could detect mumps virus RNA in approximately 70% of both throat swabs and CSF samples, while, by tissue culture, mumps virus was isolated from only approximately 20% of CSF and 50% of throat swab samples. In addition, the real-time PCR assay could be developed easily into a quantitative assay for clinical specimens containing more than 1,800 copies of mumps virus RNA/ml by using serial dilutions of the standard plasmid. The results suggest that the real-time PCR assay is useful for identification of mumps virus infections, not only in typical cases, but also in suspected cases, which show only symptoms of meningitis or encephalitis.
Latent varicella-zoster virus (VZV) has been demonstrated in the human trigeminal and thoracic ganglia by means of nucleic acid hybridization. However, the human geniculate ganglia in VZV latency have not been examined. Tissue DNA extracted from the trigeminal and geniculate ganglia of a newborn and 7 adults was examined by polymerase chain reaction with a pair of VZV-specific primers. None had symptoms of recent infection with VZV (chickenpox or shingles). VZV DNA was detected in 11 (79%) of 14 trigeminal ganglia and in 9 (69%) of 13 geniculate ganglia of the adults. VZV DNA was not detected in either type of ganglion from the newborn or from 1 adult who was seronegative for VZV antibodies. These findings indicate that VZV becomes latent in human geniculate ganglia after primary infection and suggest the possibility that reactivation of the virus from the geniculate ganglia may cause Ramsay Hunt syndrome.
Abstract. Herpesvirus cyprini (CHV) genome was traced in carp, Cyprinus carpio L., after acute infection by the method of in situ hybridization with biotinylated probes. The viral genome was detected in several tissues including cranial nerve ganglia. Subcutaneous tissue and spinal nerves. However, at this stage, viral antigens were not detected and the virus was not isolated. The viral genome was also detected in the same fish tissues when papillomas were present which contained viral antigens and even infective virus particles. After papilloma regression, the viral genome was still detected in these tissues. It is suggested that CHV becomes latently established in cranial nerve ganglia, subcutaneous tissue and spinal nerves, and is associated with the induction and recurrence of papillomas.
Nervous tissue lesions were retrospectively studied for detection of productive varicella zoster virus (VZV) infection in 33 autopsied cases, including 19 herpes zoster (HZ) (10 trigeminal, nine spinal) and 14 cases of nodular brainstem encephalitis without HZ. Immunocytochemistry for VZV antigens and in situ hybridization with a biotinylated VZV DNA probe were used on formol-fixed paraffin sections. Peripheral and central nervous system, skin and striated muscle were investigated in serial sections; available tissue blocks, however, varied between cases. Varicella zoster virus production (both antigen and DNA) in nervous tissue was found in HZ cases but only of short survival after a rash of up to 7 wks (eight out of 12 patients). Varicella zoster virus was visualized in nerve cells, glial cells, Schwann cells and blood vessels. In the central nervous system (CNS), VZV was detected in trigeminal nuclei (one out of 10 brains) or disseminated nodular brainstem lesions (one out of 10 brains), in subependymal microvessels (one out of 10 brains) or vasculitic arteries (two out of 19 brains or spinal cords). In the peripheral nervous system (PNS), VZV (DNA and antigen) was found in neurons and satellite cells of sensory ganglia (four out of seven cases with sampling of ganglia), and in damaged nerve fibres including a muscle nerve in one case; myositis with VZV in affected muscle fibres was found in the latter case. In nodular brainstem encephalitis, one case contained VZV within nodular lesions. We conclude that (i) VZV neural spread is suggested by detectable virus in ganglia, nerve fibres and CNS target nuclei; (ii) haematogenous spread of VZV is suggested by detection of virus in CNS microvessels and in disseminated brainstem encephalitis; (iii) VZV myositis may occur in zosteric myotomes; and (iv) VZV is a possible agent in nodular brainstem encephalitis.
We have used 12 restriction enzymes to analyse the DNA of 24 clinical isolates of VZV derived from 12 patients in order to compare isolates derived from different individuals and derived serially from the same individual. As reported previously, only a small proportion of the isolates differed with respect to the presence or absence of restriction sites. However, we found that the size of DNA fragments generated from all the isolates derived from different patients varied in any of four regions, one of which was first recognized in this study. In one case, where multiple isolates recovered from the same individual were analysed, each was distinguished from the others not only by differences in the variable regions but also by the presence or absence of a restriction site in a nonvariable region. This suggests that multiple strains of VZV can be present in the same human host.
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