• 제목/요약/키워드: HSV PCR

검색결과 18건 처리시간 0.039초

무균성 뇌막염과 뇌염으로 입원한 성인 환자 뇌척수액에서 중합효소 연쇄반응에 의한 HSV, VZV, HHV-6의 검출 (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)

  • 박혜경;우소연;김현진;정영해
    • 대한바이러스학회지
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    • 제30권3호
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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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중합효소 연쇄반응에 의한 수포액, 혈액과 관절액에서 단순포진 바이러스 1, 2와 대상포진 바이러스의 검출과 감별 (Detection and Differentiation of Herpes Simplex Virus 1 and 2, and Varicella-Zoster Virus in Vesicle Fluid, Joint Fluid and Serum using PCR Method)

  • 박혜경;우소연;김현진;이정화
    • 대한미생물학회지
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    • 제35권2호
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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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중합효소연쇄반응으로 확진된 Herpes Simplex virus 뇌염 1례 (A Case of Type 1 Herpes Simplex Virus Encephalitis Detected by Polymerase Chain Reaction)

  • 박대영;이준수;이영호;손영모
    • Pediatric Infection and Vaccine
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    • 제3권2호
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    • pp.207-213
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    • 1996
  • Herpes simplex virus(HSV) infections of the CNS are associated with significant morbidity and mortality even when appropriate antiviral therapy is administered. HSV infections of the brain can be subdivided into two categories : neonatal HSV infections, which usually are caused by HSV type 2, and herpes simplex encephalitis(HSE), which occur in patients over 3 months old and is nearly uniformly caused by HSV type 1. The clinical presentation of HSE is one of the focal encephalopathic process associated with altered levels of consciousness, fever, focal seizures and hemiparesis. But because of the lack of pathognomic clinical presentation and diagnostic procedure, the efforts to develop alternative diagnostic procedure have led to the use of new diagnostic technique such as polymerase chain reaction(PCR). We report a case of HSV type 1 encephalitis in 13 month old male infant who presented with altered level of consciousness, fever and focal seizures. With the use of the PCR, HSV-1 DNA was detected in cerebrospinal fluid from the patient. The symptoms and signs of encephalitis subsided by treatment with acyclovir in 14 days.

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Hydrodynamic-based Procedure를 이용한 간에서의 HSV1-tk 발현 확인을 위한 방사표지 5-(2-iodovinyl)-2'-deoxyuridine (IVDU)의 영상연구 (Imaging of Herpes Simplex Virus Type 1 Thymidine Kinase Gene Expression with Radiolabeled 5-(2-iodovinyl)-2'-deoxyuridine (IVDU) in liver by Hydrodynamic-based Procedure)

  • 송인호;이태섭;강주현;이용진;김광일;안광일;정위섭;천기정;최창운;임상무
    • Nuclear Medicine and Molecular Imaging
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    • 제43권5호
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    • pp.468-477
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    • 2009
  • 목적: Hydrodynamic-based procedure는 손쉽고 간편한 비바이러스성 유전자 전달 방법으로 특히 간특이적으로 발현하는 특징을 가진다. 단순 헤르페스 바이러스 제 1 형 티미딘 키나제(herpes simplex virus type 1 thymidine kinase, HSV1-tk)와 다양한 기질을 이용한 비침습적 HSV1-tk 유전자 영상시스템이 널리 연구되어왔다. 본 연구에서는 HSV1-tk 유전자를 hydrodynamic-based procedure를 이용하여 전달한 후, HSV1-tk의 보고 기질로 알려진 5-(2-iodovinyl)-2'-deoxyuridine (IVDU)을 이용하여 간 특이적인 HSV1-tk 유전자 발현 영상을 획득하고자 하였다. 대상 및 방법: HSV1-tk 유전자와 녹색형광유전자를 가진 각 플라스미드 벡터를 마우스에 hyodynaminc injection을 통해 전달하고, 24 시간 뒤 유전자의 발현을 확인하기 위해 RT-PCR, 생체형광영상, 핵의학영상, 전신자가방사영상 그리고 생체분포를 시행하였다. 결과: 각 플라스미드 벡터를 전달한 간으로부터 추출한 전체 RNA를 이용하여 RT-PCR을 수행한 결과, 각각 HSV1-tk유전자와 녹색형광단백 유전자의 특이적인 밴드를 관찰할 수 있었다. 생체 분포 결과, pHSV1-tk 벡터를 전달한 마우스의 간에서 특이적인 [$^{123}I$]IVDU의 섭취를 보였다. 생체형광영상에서는pEGFP-N1 벡터를 전달한 마우스의 간에서는 유의한 형광신호를 나타내었다. 전신자가방사영상과 감마카메라 영상에서 pHSV1-tk 벡터를 전달한 마우스의 간에서 방사표지 IVDU가 국소적으로 집적되는 것을 확인하였다. 결론: 본 연구에서 hydrodynamic-based procedure는 간특이적으로 플라스미드 DNA를 전달하는데 효과적이며 전달된 유전자의 발현을 분자영상학적인 방법으로 확인하였다. 따라서 Hydrodynamic injection을 통해 HSV1-tk유전자와 목적 유전자의 공동발현은 방사표지 IVDU에 의해 목적 유전자의 발현을 정량평가하는데 유용할 것으로 기대된다.

Lack of Association between Herpes Simplex Virus Type 2 Infection and Cervical Cancer - Taq Man Realtime PCR Assay Findings

  • Farivar, Taghi Naserpour;Johari, Pouran;Shafei, Shilan;Najafipour, Reza;Reza, Najafipour
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권1호
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    • pp.339-342
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    • 2012
  • Background: About one third of the human population suffer cancer during their lifetime and more than 20% of total morbidity is related to neoplasia. Cervical cancer is generally the most common cancer in developing countries and the second most common in women globally. The role of human papilloma viruses viruses in its induction is clear. However, the involvement of hepres simplex virus type 2 (HSV-2) is controversial. Therefore a survey was conducted of the prevalence of HSV-2 in patients with cervical cancer and also healthy people with sensitive and quantitative Taq Man real-time PCR assay. Materials and methods: Seventy six formaldehyde fixed paraffin embedded tissue specimens from patients with histologically proven history of cervical cancer as well as 150 control blocks were sectioned for deparaffinization and DNA extraction. Results: There was no HSV-2 DNA in our patient specimens but four control samples were positive, all with a history of hysterectomy. Conclusion: Considering the absence of any positive viral HSV-2 DNA in our patients and also the presence of four positive specimens among our controls, we did not find any relationship between the presence of HSV-2 DNA and cervical cancer.

Prevalence of Human Papillomavirus and Herpes Simplex Virus Type 2 Infection in Korean Commercial Sex Workers

  • Yun, Hae-Sun;Park, Jeong-Joo;Choi, In-Kyung;Kee, Mee-Kyung;Choi, Byeong-Sun;Kim, Sung-Soon
    • Journal of Microbiology and Biotechnology
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    • 제18권2호
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    • pp.350-354
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    • 2008
  • In order to investigate the prevalence of sexually transmitted viruses such as human papillomavirus (HPV) and herpes simplex virus (HSV) in Korean commercial sex workers (CSWs), we selected 188 CSWs (age range 20-44 years, median age 24 years) who regularly visited one public health center in Seoul, Korea. HPV genotypes were analyzed by using a HPV DNA Chip, and an enzyme-linked immunosorbent assay (ELISA) was used to detect type-specific IgG against HSV2 antibody identifying seropositivity for HSV2 infection. Polymerase chain reaction (PCR) was performed with specific primers to detect HPV and HSV1/2 in cervical swabs from the CSWs. The prevalence of HPV infection was 83.5% in 188 cervical swab specimens and the main high-risk HPV genotypes were HPV16, 18, 56, and 58. The principal low-risk HPV genotypes were HPV6 and 11. The prevalence of HSV1/2 DNA was 13.8% and HSV2 seroprevalence was 86.2%. These results suggest that high frequencies of HPV and HSV2 infection might contribute to the rapid spread of STD viruses in CSWs in Korea. Additionally, an understanding of why high-risk HPV genotypes are so prevalent could provide guidelines for prophylactic vaccine development in Korea.

Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)

  • Uh, Hong-Sun;Park, Jong-Kuk;Kang, Hyun;Kim, Soo-Young;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • 제11권3호
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    • pp.482-490
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    • 2001
  • The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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재발성 아프타성 궤양 환자의 타액에서 Herpes Simplex Virus, Varicella Zoster Virus, Helicobacter pylori 그리고 Candida 검출 (Detection of Herpes Simplex Virus, Varicella Zoster Virus, Helicobacter Pylori and Candida in Saliva of Patients with Recurrent Aphthous Ulceration)

  • 허웅;윤창륙;안종모
    • Journal of Oral Medicine and Pain
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    • 제30권3호
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    • pp.319-328
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    • 2005
  • 재발성 아프타성 궤양 환자의 타액에서 구강내 궤양성 병소를 유발할 수 있고 감염성이 비교적 높은 미생물로 알려진 Herpes Simplex virus, Varicella Zoster virus, Helicobacter pylori 그리고 Candida가 발현되는지 여부와 병소의 발생과 상관관계가 있는지를 알아보고자 조선대학교 치과병원 구강내과에 내원한 재발성 아프타성 궤양을 가진 환자 29명과 대조군 29명의 타액을 이용하여 PCR과 배양을 통해 발현율을 비교한바 다음과 같은 결과를 얻었다. 1. HSV DNA는 재발성 아프타성 궤양 환자군에서 41.4%, 대조군에서 55.2%가 발현되었으나 두 군간에 유의한 차이는 없었고(P>0.05), VZV DNA는 두 군에서 모두 나타나지 않았다. 2. H. pylori DNA는 재발성 아프타성 궤양 환자군에서 27.6%, 대조군에서 48.3%가 발현되었으나 두 군간에 유의한 차이는 없었다(P>0.05). 3. Candida는 재발성 아프타성 궤양 환자군에서 13.8%, 대조군에서 6.9%가 배양되었으나 두 군간에 유의한 차이는 없었다(P>0.05). 이상의 연구를 종합하여 보았을 때, HSV, VZV, H. pylori 그리고 Candida는 재발성 아프타성 궤양의 발생에 직접적인 역할을 한다고는 볼 수 없으므로 향후 더 많은 표본을 대상으로 다른 미생물이 병소 발생의 유발요인으로 작용하는지 연구하여야 할 것으로 사료된다.

엔테로바이러스 검출을 위한 real-time nucleic acid sequence-based amplification (NASBA), reverse transcription-PCR (RT-PCR) 및 바이러스 배양법의 비교 (Comparison of the Real-Time Nucleic Acid Sequence-Based Amplification (NASBA) Assay, Reverse Transcription-PCR (RT-PCR) and Virus Isolation for the Detection of Enterovirus RNA.)

  • 나영란;조현철;이영숙;빈재훈;최홍식;민상기
    • 생명과학회지
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    • 제18권3호
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    • pp.374-380
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    • 2008
  • 본 연구는 무균성수막염 의심환자의 다양한 검체로부터 enterovirus의 진단을 위하여 real-time NASBA, 2 step RT-PCR 시험과 세포배양 시험을 각각 실시하여 각 시험법의 검출율, 특이도, 사용자 편리성, 시험소요 시간, 교차오염의 가능성 등을 비교 검토하였다. 비교시험 결과 전체 292건의 검체로부터 real-time NASBA에서 145건, 세포배양에서 101건, 2 step RT-PCR에서 86건이 양성으로 나타나 real-time NASBA가 가장 검출율이 높은 시험법임을 알 수 있었다. Enterovirus 외의 무균성수막염 원인바이러스에 대한 특이도 비교 시험결과 2 step RT-PCR 시험에서 rhinovirus 10건 중 1건이 위양성 반응을 나타내어 다른 시험법에 비해 특이도가 떨어지는 것으로 나타났다. Real-time NASBA는 하나의 튜브에서 증폭과 검출이 동시에 일어나 다른 시험과 비교하여 교차오염의 가능성이 낮으며 또한 시험 소요시간이 5시간 정도로 세포배양(5-14일 소요) 및 2 step RT-PCR(9시간소요) 에 비하여 신속하게 진단할 수 있어 일선병원이나 실험실에서 enterovirus를 검출을 위하여 적용할 수 있을 것으로 사료된다.

베체트병의 동물모형에 대한 사심탕류 투여 효과에 관한 연구 (Experimental Studies on the Kinds of Sasim-tang In Behcet's Disease Symptoms in ICR Mice)

  • 이선구;안규석
    • 동의생리병리학회지
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    • 제18권4호
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    • pp.1061-1070
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    • 2004
  • Chronic oral aphthae, recurrent ulcer and uveitis are the three main festations of Behcet's disease(BD). The aetiopathogenesis of Behcet's disease is still obscure, but herpes simplex virus is one of the possible casual factors. Gamchosasim-tang (Gancaoxiexin-tang), Banhasasim-tang(Banxiaxiexin-tang) and Saenggangsasim-tang( Shengjiangxiexin-tang) are traditional medication in Oriental medicine, that has been used to treat inflammatory disease. Especially, Gamchosasim-tang used to treat Behcet's disease like symptoms. ICR mice were used for this study. The earlobe of the mice were scratched with a needle, then inoculation with 1.0×10/sup 6/ plaque forming units/㎖ of HSV type I. Virus inoculation was performed twice with 10 day interval, followed by 16 weeks of observation. Using the HSV-induced Behcet's disease mouse model, kinds of Sasim-tang were administered variously before and after inoculation. In order to. classify the symptomatic mice as having Behcet's disease like symptoms. We followed the revised Japanese classification with minor modifications. Ulceration of the mice were monitored. In addition, spleen cytokine expression were measured by polymerase chain reaction, ELISA. HSV DNA was detected in HSV inoculation mice. HSV-induced mice treated with kinds of Sasim-tang showed improvement in symptom. In RT-PCR results, IFN-γ was expressed for all groups, IL-2 was expressed for the treated groups, and IL-10 was also expressed. IL-4 was expressed nothing. In ELISA, IL-2 was increased for GSST 2, BSST 2, GSST 2, GSST3 and INF-γ was increased for GSST 2, BSST 2, SSST 2, SSST 3. This model suggest the possible role of immune response to viral infection in the development and activation of Behcet's disease.