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Shell Vail Culture를 이용한 단순포진 바이러스 분리
최태열,이규택,박해일,강정옥 대한진단검사의학회 2003 Annals of Laboratory Medicine Vol.23 No.5
목적 : 세포배양을 이용한 Herpes simplex virus (HSV)의 진단은 가장 좋은 방법이기는 하지만, 일부 검체에서는 세포병리효과(cytophathic effect, CPE)를 관찰하기까지 많은 시간이 요구된다. 이에 연구자들은 HSV의 진단을 위하여 신속하고 예민한 배양 방법을 개발하였다.방법 : HSV 감염이 의심되는 13명의 환자에서 채취된 검체를연구에 사용하였다. Shell vial 내의 Vero 단층세포에 검체를 원침하여 접종한 후 배양하였으며, CPE는 도립-위상차 현미경으로5일간 매일 관찰하였다. 검체 접종 2일만에 HSV에 특이한 단클론항체를 이용하여 직접면역형광염색(direct immunofluorescencestain, DIF)을실시하였으며, DIF 염색에 음성인 검체는 shell vial내에 단층세포을 분쇄하여 새로운 shell vial에 재접종을 실시하였다. 중합효소연쇄반응(polymerase chanin reaction, PCR)법은모든 검체에서 실시하였다.결과 : CPE는 접종 후 1, 2, 3, 4 및 5일에 각기 30 (64%), 39(83%), 43 (92%), 44 (94%) 및 46 (98%)예에서 관찰되었다.DIF염색에서는 접종 2일 만에 46 (98%)예가 양성으로 발견되었다. DIF 음성인 검체를 재 접종한 결과 1일 만에 새로운 7예에서328 최태열이규택박해일 외 1인CPE를 관찰할 수가 있었다. PCR법에서 47 (100%)예가 양성으로 나타났다.
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단순포진 바이러스 검출 및 형결정을 위한 간접면역형광법과 Nested Multiplex-중합효소연쇄반응의 비교
최태열,안유헌 대한진단검사의학회 2004 Annals of Laboratory Medicine Vol.24 No.2
Background : Herpes simplex virus type 1 (HSV-1) is linked specifically to oral and lip infections while HSV-2 is involved in genital infections. We evaluated a recently reported nested-multiplex ploymerase chain reaction (NM-PCR) for the detection and typing of HSV and compared the results with indirect immunofluorescence (IF) after cell culture. Methods : One hundred thirty three specimens were received from patients suspected of having clinical HSV infections. HSV was cultured by the shell vial method and stained with type specific monoclonal antibodies. NM-PCR was performed using crude samples. Results : HSV was detected in 45 (33.8%) and 46 (34.6%) of the 133 specimens by IF and NMPCR, respectively. All of the HSV IF positive specimens were also positive by NM-PCR. Typing by the two methods concurred in all but two of the 45 specimens; the two specimens were typed as HSV-1 and HSN-2, respectivey, by IF, and both as HSV-1 and HSV-2 by NM-PCR. Conclusions : Our results suggest that NM-PCR is a rapid and sensitive method for the detection and typing of HSV. (Korean J Lab Med 2004; 24: 113-8) 목적 : HSV-1은 주로 구강 및 구순에 감염을 일으키고, HSV-2는 생식기에 감염을 일으킨다. 연구자들은 HSV의 검출 및 형결정을 위한 Coyle 등의 nested multipex-polymerase chain reac-tion(NM-PCR) 방법을 평가하고자 이 연구를 실시하였으며, 세포배양 후 간접면역형광법의 결과와 비교 평가하였다. 방법 : HSV 감염이 의심되는 133 검체를 검사 대상으로 하였다. 검체는 shel vial을 이용한 세포배양 후 HSV-1 및 HSV-2 단클론항체를 이용하여 간접면역형광 염색을 실시하였다. 원래 검체는 HSV 검출 및 형결정 검사를 위하여 NM-PCR (J Virol Me -thods 1999; 83: 75-82)을 실시하였다. 결과 : 세포배양 후의 간접면역형광법은 45/133 (33.8%)예,NM-PCR은 46/133 (34.6%)예에서 HSV를 검출하였다. 간접면역형광법에서 HSV 양성인 검체는 모두 NM-PCR에서 양성이었다. 두 방법간에서 형결정 일치율은 45 검체 중 43 검체로 95.6%이었다. 나머지 2 검체는 간접면역형광법에서 각기 HSV-1과 HSV-2이었으나 NM-PCR에서는 HSV-1과 HSV-2 모두가 각 검체에서 동정되었다. 결론 : NM-PCR은 HSV감염 검출 및 형결정에 신속 정확한 검사방법이다.
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최태열 한양대학교 의과대학 1997 한양의대 학술지 Vol.17 No.2
Detection of HIV antibodies is still the most efficient and most common way to determine whether an individual has been exposed to HIV and to screen blood and blood products for this infections agent. A test is considered positive when assay such as the enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and western blots (immunoblots)are considered reactive. A positive test result indicates exposure and, outside of the perinatal and neonatal period, is presumed to indicate infection by teh virus. The ELISA is the most commonly used to screen for HIV because of its relatively low cost, standardized procedure, high reliability, and ability to give rapid result. Under optimal laboratory conditions, the sensitivity and specificity in most cases are>99%. The Western blot assay is the test most commonly used for confirming the presence of HIV-specific antibodies. However, western blots are more expensive and more time consuming than ELISAs, and they require more technical expertise owing to subjective interpretations, Specificity and sensitivity are high but are some what dependent on the interpretive criteria employed. A definitive test for an active infection is the recovery of HIV from cells or cell-free fluids of the infected individual by using the solid-phase capture technique, indirect immunofluorescence assay, ploymerase chain reaction... etc. Surrogate markers (CD4+ -CEll count, P24 antigen in plasma, 2-microglobulin... etc.) are not only important in predicting the out come of HIV infection but also useful in determining when treatment should be administered. Inconclusion, while awaiting the development of more-effective antiviral therapies, early testing for HIV in potentially infected individuals is recommended to allow prompt medical treatment and surveillance and to prevent the spread of the virus to other individuals.
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최태열,강정옥,배현주 대한임상미생물학회 2011 Annals of clinical microbiology Vol.14 No.1
Background: We investigated whether culture using an automated blood culture system enhances the recovery of bacteria and fungi from body fluids other than blood when compared to conventional solid media culture methods. Methods: A total of 734 specimens [ascites (n=457), bile (n=5), CAPD (n=28), CSF (n=32), joint fluids (n=165), pericardial fluid (n=17), and pleural fluid (n=30)] were included in the study. Half of the volume of each specimen was inoculated directly into automated blood culture bottles (bioMeriux, Marcy-I’Etoile, France). The remaining volume was inoculated onto conventional solid media (sheep blood agar, chocolate agar, and phenylethyl alcohol agar) after centrifuging at 3,000 rpm for 10 min. Results: Clinically significant microorganisms were isolated from 62 specimens (8.5%) by automated blood culture and 61 specimens (8.3%) by the conventional solid media culture (kappa index: 0.81, 95% confidence interval: 0.75∼0.89). Contamination was observed in 11 (1.8%) of the automated blood culture specimens and 3 (0.4%) of the solid media culture specimens. The mean turnaround times of the automated blood cultures and the conventional solid media cultures were 3.7 and 2.8 days, respectively (P< 0.0001). Conclusion: Compared with conventional culture methods, no improvement in the recovery of clinically significant microorganisms was noted with the use of the automated blood culture system for the culture of body fluids other than blood.
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