• Title/Summary/Keyword: DNA typing

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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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Clinical aspects of an outbreak of Serratia marcescens infections in neonates (신생아 중환자실에서 Serratia marcescens에 의한 집단 감염 발생에 대한 고찰)

  • Sung, Min-Jung;Chang, Chul-Hun;Yoon, Yeon-Kyong;Park, Su-Eun
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.500-506
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    • 2006
  • Purpose : We evaluated an outbreak of Serratia marcescens infections in 24 neonates in a neonatal intensive care unit(NICU). Methods : From January to August, 2004 a nosocomial outbreak of S. marcescens occurred in our NICU. We describe the clinical characteristics of the outbreak and analyse the risk factors for infections with S. marcescens. After the outbreak stopped, 7 isolates from blood were typed using rapid amplified polymorphic DNA analysis(RAPD). Results : S. marcescens was isolated from 24 neonates, 19 infected and 5 colonized. Seven out of nineteen neonates had bacteremia, 4 had ventilator associated pneumonia, 4 had purulent conjunctivitis, 2 had UTI, 1 had meningitis and 1 had a wound infection. Three neonates died due to S. marcescens infection, 2 of 3 had ventilator associated pneumonia, 1 had meningitis complicated with abscess. The mortality rate of S. marcescens infection was 15.8%. Factors associated with S. marcescens infections were previous antibiotic therapy, indwelling catheter and use of ventilators. The isolated strains were resistant to most antibiotics, but frequently sensitive to imipenem, bactrim and amikacin. RAPD typing results show that at least 3 epidemic strains were related with this outbreak. But one genotype was predominant type in this outbreak. The control measures were instituted and the outbreak stopped within 2 months. Conclusion : S. marcescens can cause rapidly spreading outbreaks associated with fatal infections in neonates. If S. marcescens is isolated from clinical specimens, meticulous infection control measures and epidemiologic investigations should be done at an early stage of the outbreak.