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중합효소연쇄반응과 제한효소분석을 이용한 조갑진균증 원인 진균의 진단
채희재,백승철,조백기 대한의진균학회 1999 대한의진균학회지 Vol.4 No.1
Background: Onychomycosis has become one of the common fungal infection. However, highly reliable and sensitive methods of detecting and identifying causative fungi of onychomycosis are not established yet. Polymerase chain reaction (PCR) analysis of clinical specimens including blood, sputum, urine, and cerebrospinal fluid collected from patient systemically infected fungus is known as a sensitive diagnostic method. But it has been questionable whether PCR analysis is also applicable to onychomycosis. Objective: The purpose of this study was to develop a DNA-based diagnostic method to improve the sensitivity and specificity of detection and identification of pathogenic fungi of onychomycosis. Methods: To detect the fungi in the nail, PCR was performed by using 4 sets of primer (TR1 & TR2, NS5 & NS6, B2F & B4R and CA1 & CA2) designed in conserved sequences of the small ribosomal subunit (185-rRNA) genes and restriction enzyme analysis of amplified product by Hae Ⅲ was done to identify species. Nail specimens were obtained from 19 cases of onychomycosis confirm by fungus culture. Results: 1. Preparation of nail powder, which is necessary for removal of keratin, and composition of Iysis buffer with guanidinium thiocyanate, Tris-HCI, and β-mercaptoethanol are the most proper modalities for isolation of fungal DNA from fungus-infesting nails. 2. Specific fragments of the 18S-rRNA gene of fungi, 581 bp, 308 bp, 688 bp and 1106 bp were amplified respectively. From sequences of 18S-rRNA gene of fungi by universal primers, dermatophytes (Trichophyton rubrum, Trichophyton mentagrophytes) and yeast (Candida albicans) yielded identical products. 3. Using Hae Ⅲ endonuclease, digested patterns of fragment of Trichophyton rubrum and Candida albicans resulted in different pattern. Conclusion: This method released enough DNA from fungus-infected nails to result in proper amplification and it can be possible to differentiate dermatophytes, yeasts, and molds using Hae Ⅲ endonuclease. The present study is the first one to demonstrate the feasibility of this molecular biologic approach to identify fungi in the infected nail. Therefore, precise detection and identification of the causative fungi would be of help in investigating distribution of the causative fungi of onychomycosis as well as appropriate treatment of the disease.
10 년간 경상의대 의학과 학생의 A 형 간염 항체 양성률 변화와 경상의대 소아과 근무 의료인의 항체 양성률
임재영,고경혁,조명제,이시은,조중현,윤희상,백승철,이우곤,이광호,우향옥,조윤경,박찬후,정양숙,이수진 대한소화기학회 1999 대한소화기학회지 Vol.33 No.4
Background/Aims: Korean population has been suggested to be minimally exposed to hepatitis A virus (HAV) for recent 20 years. Therefore, hepatitis A epidemics may occur among medical students and medical personnels who care the asymptomatic patients with hepatitis A. We investigated the changes in anti-HAV IgG positive rate among the medical students for 10 years and the curren anti-HAV IgG positive rate among the medical personnels in Chinju. Methods: Serum anti-HAV IgG was measured using the commercially available HAVAB radioimmunoassay kit. The sera had been collected from 780 sophomore medical students during 1988-1997 and from 95 pediatric medical personnels at the Gyeongsang National University Hospital in December, 1997. Results: The anti-HAV IgG positive rate of the sophomore medical students was 87.5% in 1988, 86.7% in 1989 83.8% in 1990, 75% in 1991, 85% in 1992, 70% in 1993, 56.3% in 1994, 58.8% in 1995, 53.8% in 1996 and 38.8% in 1997. The anti-HAV IgG positive rate among the pediatric medical personnel (mean age ±S.D.; 26.4 ±3.1) was 65.3%. Conclusions: These suggest that the population susceptible to HAV exists among medical students and pediatric medical personnels. Therefore, we should se up the preventive modalities against hepatitis A virus infection among these high risk groups.
영아에서 Helicobacter pylori 감염에 대한 항체반응
임재영,고경혁,조명제,김은아,김윤옥,윤희상,백승철,이우곤,이광호,우향옥,박찬후,정양숙 대한소화기학회 2000 대한소화기학회지 Vol.35 No.6
Background/Aims: The early stage of Helicobacter pylori (H. pylori) infection may show only anti-H. pylori IgM antibody response in young infants. We tried to find out the antigens of H. pylori which stimulate antibody response during the early stage of infection using IgM, IgA, and IgG immunoblot analyses. Methods: Sera of 48 infants were collected at birth and 7 months of age under the permission of parents and sera of 17 infants were also collected at 3 months and 9 months after birth. For those sera, IgM, IgA, and IgG immunoblotting were performed. Results: At the first study, seventeen of 48 (35.4%) infants show 120 kDa band on IgM or IgA immunoblot at 7 months of age. Four infants showed 120 kDa band on IgM immunoblot but not on IgA immunoblot. They were assumed to be infected very recently. At the second study, 2 of 17 showed strong IgM immune response to H. pylori at 3 months of age. The 120, 61, 60, 56, 54, 35, 29, 25, 18 kDa bands were recognized on IgM immunoblot in both studies. Conclusions: At 7 months of age, 35.4 % of infants were infected with H. pylori. Antigens of H. pylori which stimulate antibody response during the early stage of human infection were 120, 61, 60, 56, 54, 47, 29, 25, 18 kDa.
소아 위생검 조직절편을 이용한 H . pylori 감염 진단에 있어서 PCR 적용의 한계
임재영,고경혁,조명제,김윤옥,오영균,박철근,백승철,이우곤,이광호,우향옥,최명범,조윤경,정양숙,박찬후,윤희상,맹국영 대한소화기학회 1998 대한소화기학회지 Vol.31 No.1
Background/Aims: We tried to evaluate the clinical usefulness of the PCR for the diagnosis of H. pylori infection in children and to identify the possible false positive results by the PCR due to remaining H. pylori in the working channel of an endoscope or in the biopsy forceps. Methods: Forty seven urease test samples with three gastric biopsy specimens, 6 collections of 15 ml flushing distilled water after the end of working channel disinfection, 11 15 ml-distilled-water batches as the negative controls, and one H. pylori positive paraffin block as the positive control were collected at the Gyeongsang National University Hospital. The Hel-2 primer set (GTGTGCGGGCTTACAAGGAT, CGTTAGCGTTCATCACACTC) and a 34 cycle amplification were used Results: All of the seventeen specimens of urease tested positive within 6 hours and the ten specimens of urease also tested positive within 48 hours were PCR positive. Eighteen of the 20 specimens of urease tested negative and were also PCR positive. Three of the 6 specimens of 15 ml flushing distilled water were found to be PCR positive. All the negative controls were PCR negative and the one positive control was PCR positive. Conclusions: The clinical usefulness of PCR using gastric biopsy specimens in children was limited due to the possible dead or live H. pylori remaining in the biopsy channel.