KISEP Head and Neck Korean J Otolaryngol 2001;44:1171-6 만성편도염에서의 Epstein-Barr Virus 의검출률과유전자형분포 최영철 1 이숙경 2 권용순 2 강진한 3 박용수 1 전은주 1 Detection Rates of Epstein-Barr Virus from the Chronic Tonsillitis and Its Typing Young-Chul Choi, MD 1, Suk Kyeong Lee, PhD 2, Yong Soon Kwon 2, Jin-Han Kang, MD 3, Yong-Soo Park, MD 1 and Eun-Ju Jeon, MD 1 1 Department of Otolaryngology-Head and Neck Surgery, 2 Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, and 3 Pediatrics, College of Medicine, The Catholic University of Korea, Seoul, Korea ABSTRACT Background and ObjectivesEpstein-Barr virus EBV is a DNA virus and a type of Herpes virus. Two different types of EBV exist based on the DNA sequence divergence of the EBV genome, and several differences exist between the two types. There are no reports about EBV detection rates and typings associated in chronic tonsillitis, despite the known fact that tonsil is one of the most common sites EBV resides. The aims of this study were to investigate the prevalence of EBV infection in tonsils of children and adults as well as to compare the detection rates and the distribution of types of EBV in tonsillitis patients with normal controls. Materials and MethodsPalatine tonsil tissues were obtained from seventy adults 20 normal controls and 50 chronic tonsillitis and one hundred children. Using polymerase chain reaction PCR, EBV genome was detected and typing was performed. We confirmed EBV by Southern blot hybridization and in situ hybridization. ResultsThe detection rates of EBV in chronic tonsillitis of adults were higher than those in children with idiopathic tonsillar hypertrophyith and normal controls. The detection rates of type 1 EBV showed no differences among the four groups. The detection rates of type 2 EBV in ITH and chronic tonsillitis of adults and children were higher than those in the normal controls. Conclusion Our results suggest that tonsillitis is caused by EBV and that the type 2 EBV plays a more important role in tonsillitis. Korean J Otolaryngol 2001;44:1171-6 KEY WORDSEpstein-Barr virus Palatine tonsil Typing. 1171
DNA의추출 1172 Tabel 1. Primer sequences for EBV detection Sequences 1st sense 5 -AAGGAGGGTGGTTTGGAAAG-3 1st antisense 5 -AGACAATGGACTCCCTTAG-3 2nd sense 5 -ATCGTGGTCAAGGAGGTTCCA-3 2nd antisense 5 -ACTCAATGGTGTAAGACGAC-3 EBV 검출을위한중합효소연쇄반응 (PCR) 중합효소연쇄반응 (PCR) 을통한 EBV의감염형의분류 Korean J Otolaryngol 2001;44:1171-6
Southern hybridization In situ hybridization 통계처리 Table 2. Detection rate of EBV Group Detection rate Cases *ITH 50 25/50 Chronic tonsillitis in children 62 31/50 Chronic tonsillitis in adults 72 36/50 Healthy adults 45 9/20 ITHIdiopathic Tonsillar Hypertrophy, p0.05 P 1 2 3 4 5 6 7 8 9 10 M 11 12 13 14 15 16 17 18 19 20 21 22 N Fig. 1. EBV detection by nested PCR. PCR products of palatine tonsil DNA was reamplified with nested primer sets and separated on 2 agarose gel. A band of 208 bp indicates the presence of EBV DNA. P-globin, Nnegative control distilled water, Mmarker. 1173
Table 3. Typing of EBV from palatine tonsils of healthy adults and tonsillitis patients Group Detection rate Cases Type 1 Type 2 Type 1 2 *ITH 24 12/50 8 4/50 18 9/50 Chronic tonsillitis in children 22 11/50 8 4/50 28 14/50 Chronic tonsillitis in adults 30 15/50 6 3/50 30 15/50 Healthy adults 35 7/20 0 0/20 10 2/20 ITHIdiopathic Tonsillar Hypertrophy Fig. 3. In Situ Hybridization. Positive signal black arrow of Not 1, Pst 1 probe hybridization was seen on the hypernuclear, large lymphocyte of lymphoproliferative paracortical area 100. 1 2 3 4 5 6 7 8 9 10 11 12 13 P2 P1 N Fig. 2. Southern blot hybridization. For EBV detection we used a 3 end-labelled oligonucleotide probe specific to EBNA-3C sequence. P1positive control of type 1 Namalwa cell, P2positive control of type 2 SNU-99 cell, Nnegative control distilled water. 1174 Korean J Otolaryngol 2001;44:1171-6
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