• Title/Summary/Keyword: VZV nested PCR

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Detection of Coinfection and Persistent Infection of Adenovirus and Varicella-Zoster Virus in Synovial Fluids from Synovitis Patients by Nested-PCR (Nested 중합효소연쇄반응을 이용하여 활막염 환자의 관절액으로부터 아데노바이러스와 대상포진바이러스의 이중감염과 지속감염의 검출)

  • Park, Hae-Kyung;Woo, So-Youn;Kim, Hyun-Jin
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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    • pp.179-187
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    • 2000
  • The etiology of rheumatic arthritis (RA) is associated with a number of genetic and environmental factors, but is not definitively elucidated. Recently, more attention has been paid to the possibility of microbial etiology in the pathogenesis of RA, because many different infectious agents have been reported to precede the onset or exacerbation of RA. Adenovirus (ADV) may be one cause of persistent or recurrent inflammatory arthritis. Varicella zoster virus (VZV) arthritis is detected frequently in RA patients treated with low dose methotrexate. The demonstration of simultaneous presence of both viral agents of specific viral nucleic acid in synovial fluids from synovitis patients would provide more direct evidence for arthritis etiological relationship, but there are no confirmed results. Therefore, we studied the ability of adenovirus and VZV to establish coinfection and persistent infection in synovial fluid from synovitis patients. The presence of viral agents in the synovial fluid demonstrated by isolation of cell culture, enzyme immunoassay and nested-PCR. The synovial fluids were also investgated for the presence of viral nucleic acid by nested-PCR using specific primer. ADV produced 220 bp and VZV produced 447 bp by each nested-PCR with specific primers. We detected 4/6 cases (66.7%) with persistent infection of ADV and 5/6 cases (83.3%) of VZV with 13 synovial fluids (between 7 to 52 day intervals) from synovitis patients by monoclonal ErA and nested-PCR. 21/28 cases (75%) with coinfection of adenovirus and VZV with synovial fluids from synovitis patients by nested-PCR. ADV and VZV coinfection and persistent infection of synovial fluids may provide a chronic antigenic stimuli to the immune system therefore provoking a continuing inflammatory response and caused the possibility of synovitis and arthritis.

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Detection of Herpes Simplex Virus, Varicella-Zoster Virus and Human Herpes Virus-6 by PCR in Cerebrospinal Fluid from Hospitalized Adult Patients with Aseptic Meningitis or Encephalitis (무균성 뇌막염과 뇌염으로 입원한 성인 환자 뇌척수액에서 중합효소 연쇄반응에 의한 HSV, VZV, HHV-6의 검출)

  • Park, Hae-Kyung;Woo, So-Youn;Kim, Hyun-Jin;Chong, Young-Hae
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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    • pp.171-178
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    • 2000
  • Herpes simplex virus, Varicella zoster virus and Human herpes virus-6 caused central nervous system infections and latent infections but there is no data of the 3 viruses being tested from the same cerebrospinal fluid samples with aseptic meningitis or encephalitis in adults patients. These viruses produced similar neurologic symptoms but difficulties existed in differentiating of etiologic agents and therefore the viruses needed to be detected in the early state. Herpes simplex virus encephalitis (HSVE) in adults, if not treated promptly was fatal. If treated with antiviral drugs in the early phase of encephalitis, neurologic sequales decreased by 65%. Recently, a PCR method for detection of HSVE with CSF was developed. VZV primary and secondary infections caused neurologic symptoms of encephalitis or meningitis. The second frequency of adult encephalitis that caused VZV were reported. HHV-6 caused CNS latent infection that was studied with normal adults brains. But there is no data of HSV, VZV and HHV-6 for aseptic meningitis and encephalitis of Korean adults through etiologic study. We cultured CSFs on HEp-2 cells and simultaneously tested for HSV PCR, VZV nested PCR and HHV-6 PCR with 8 specific primers. The PCR results of CSF from meningitis Korean adults were 13/19 (68.4%) for HSV, 10/19 (52.6%) for VZV and 12/19 (63.2%) for HHV-67/19 (36.8%) cases were triple infected HSV PCR, VZV PCR and HHV-6 PCR positive; 3/19 (15.8%) cases were dual infected HSV PCR and HHV-6 PCR positive; 1119 (0.5%) cases was VZV PCR positive. Strong viral DNA amplification of CSF means a causative virus may be present in aseptic meningitis or encephalitis patients and may cause clinical neurologic symptoms. HSV and HHV-6 viruses detection rate were higher than VZV by PCR with CSFs.

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Detection and Differentiation of Herpes Simplex Virus 1 and 2, and Varicella-Zoster Virus in Vesicle Fluid, Joint Fluid and Serum using PCR Method (중합효소 연쇄반응에 의한 수포액, 혈액과 관절액에서 단순포진 바이러스 1, 2와 대상포진 바이러스의 검출과 감별)

  • Park, Hae-Kyung;Woo, So-Youn;Kim, Hyun-Jin;Lee, Chung-Hwa
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.191-201
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    • 2000
  • The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with THP-1 and HSV-2 standard strain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of THP-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and THP-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, Band BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum form respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 47.6%) HSV was detected singly or mixed infection and 19 of those cases were HSV-2 and 1 case was THP-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, Band BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.

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