Fig.1 Procedures of the spot tests for the detection of serum HBV-DNA
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1 Key words: HBV DNA, HBV carrier, HBeAg
2 Fig.1 Procedures of the spot tests for the detection of serum HBV-DNA
3 昭 和61年2月20日 87 Table 1 Comparison of HBeAg/anti-HBe Fig.2 Comparison of the sensitivity of the spot and serum HBV-DNA Fig.3 HBeAg,anti-HBe,HBV-DNA,and tests an Line A:Reference series;al,a5:negative reference;a2,a6:positive reference Line B:HBV-DNA positive sample Line C:HBV-DNA negative sample Sera:Al,A2,B3-B8,C3-C8 Extracted sera:a5,a6,b1,b2,c1,c2 Spot test with 0.1% NaN3:B2,B6,B7,B8,C2, C6,C7,C8 Spot test without 0.1% NaN3:Al,A2,A5,A6, B1,B3,B4,B5,C1,C3,C4,C5 Volume of the serum:10u1=b3,b6,c3,c6; 30u1=B4,B7,C4,C7; 50u1=A1,A2,B5,B8,C5,C8 Volume of the extract:50u1=a5,a6,b1,b2 :HBeAg, :anti-hbe, :GPT(serum HBV glutamic-pyrubic 43),GPT異 変 動 とGPT値 そ の う ち1例 と の 関 連 に つ い て も検 討 を し た. で はFig.3に の%inhibitionが90%以 示 し た よ う にanti-HBe 持 続 し て 陽 性 で あ り,さ ら にGPTの 値 が 正 常 化 し た 後 も 検 出 さ れ た.そ 他 の2例 に つ い て は 図 示 し て い な い が,anti-HBe の 出 現 か ら2 3年 後 に 血 中HBV-DNAは れ な く な っ た.GPT値 の 検 出さ の 変 動 と血 中HBV-DNA anti-hbe出 持 続 して 正 常 値 を 示 し の 血 清43検 体 と 異 常 値 を 示 し た44例 44検 体 に お け る 血 中HBV-DNAの て 検 討 し た(Table DNAが 検 出 さ れ た.GPT異 DNAが 多 く検 出 さ れ た が,x2検 4)HBsAg消 2).GPT正 追 跡 調 査 中 にHBsAgの ア ー22例(anti-HBs陽 消 失 前(6ヵ 常 値 群 に 血 中HBV定で有意 差 はみ の血清 anti-hbs出 れ,さ 消 失 し たHBVキ 性12例)に 月 か ら24ヵ 月)お 月 か ら36カ,月)の よ び 消 失 後(6カ 3).た 体 の血 中 だ1例 にお 失 前 後 と も に 血 中HBV-DNAが の 症 例 は,Fig.4に 現 後 に もHBeAgが 示 した 如 く 持 続 して 検 出 さ 間 経 過 後 の 検 体 か ら もHBV-DNA が 検 出 さ れ た. 考 HBV感 ャ リ お け るHBsAg 血 清 そ れ ぞ れ22検 検 討 し た(Table ら に2年 検 出率 に つ い 常 値 群 は11.6%(5/ 血 中HBV- 失 例 に お け る 血 中HBV-DNA 検 出 さ れ た.こ 間 以 上 経 過 し たHBV キ ャ リ ア ー の う ち,GPTが た43例 常 値 群 は25.0%(11/44)に い て,HBsAg消 との相 関 性 現 後2年 transaminase) HBV-DNAを の 相 関 は み ら れ な か っ た. 3)transaminase値 in ら れ な か っ た, 上 で あ った に もか か わ ら ず,血 中HBV-DNAは GPT carrier 察 染 性 の 良 き 指 標 と さ れ て い るHBeAg/
4 Fig.4 One case in which persistence of HBeAg was observed after the clearance of HBsAg Table 2 Presence of serum HBV-DNA in HBV carriers Not significant(chi-square test)
5 1) Bonino, F., Hoyer, B., Nelson, J., Engle, R., Verme, G., Gerin, J.: Hepatitis B virus DNA in the serum of HBs Ag carriers: A marker of active hepatitis B virus replication in the liver. Hepatology, 1: , ) Weller, I. V. D., Fowler, M. J. F., Monjardino, J. and Thomas, H. C.: The detection of HBV- DNA in serum by molecular hybridization: A more sensitive method for the detection of complete HBV particles. J. Med. Virol., 9: 273 3) Scotto, J., Hadchouel, M., Hery, C., Yuart, J., Tiollais, P. & Brechot, C.: Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: Comparison with results for other viral markers. Hepatology, 3: , ) Lieberman, H. M., LaBrecque, D. R., Kew, M. C., Hadziyannis, S. J. & Shafritz, D. A.: Detection of hepatitis B virus DNA directly in human serum by a simplified molecular hybridization test: Comparison to HBeAg/anti-HBe status in HBsAg carriers. Hepatology, 3: , ) Tur-Kaspa, R., Keshet, E., Eliakim, M. & Shouval, D.: Detection and characterization of hepatitis B virus DNA in serum of HBe antigen-negative HBsAg carriers. J. Med.
6 Virol., 14: 17-26, ) Berninger, M., Hammer, M., Hoyer, B. & Gerin, J.L.: An assay for the detection of the DNA genome of hepatitis B virus in serum. J. Med. Virol., 9: 57-68, ) Brechot, C., Hadchouel, M., Scotto, J., Degos, F., Charnay, P., Trepo, C. and Tiollais, P.: Detection of hepatitis B virus DNA in liver and serum: A direct appraisal of the chronic carrier state. Lancet, ii: , ) Kam, W., Rall, L. B., Smuckler, E. A., Schmid, R. & Rutter, W.J.: Hepatitis B viral DNA in liver and serum of asymptomatic carriers. Proc. Natl. Acad. Sci. U.S.A., 79: , ) Tamura, I., Shimase, K., Ichimura, H., Kurimura, O., Kaneto, E., Tsuchie, H. & Kurimura, T.: Persistence of hepatitis B e antigen for a long period after the clearance of hepatitis B surface antigen. Acta Virol.,(in press). Detection of HBV DNA in the Sera of HBV Carriers by Spot Hybridization Test Ikuo TAMURA,Hiroshi ICHIMURA & Osamu KURIMURA Institute of Clinical Research,Kure National Hospital Hideaki TSUCHIE & Takashi KURIMURA Department of Virology,Tottori University School of Medicine Hepatitis B virus DNA(HBV DNA)was determined in the sera of 141 HBV carriers by spot hybridization test with cloned HBV DNA probe.the following results were obtained. 1)Spotting 10,u1 of serum directly onto nitrocellulose filter gave higher sensitivity than spotting nucleic acid extracted from 300,u1 of serum. 2)Prevalence of serum HBV DNA of HBV carriers before and after seroconversion from HBeAg to anti-hbe was studied.serum HBV DNA was detected in 84.4%(27/32)of HBeAg positive phase sera, 5.0%(1/20)of samples negative for both HBeAg and anti-hbe and 12.5%(3/24)of anti-hbe positive phase sera. 3) Of anti-hbe positive HBV carriers, serum HBV DNA was detected in 11.6%(5/43)of group consists of normal transaminase level,and 25.0%(11/44)of abnormal transaminase level.there was no significant difference in the rate of detection of serum HBV DNA between normal and abnormal carriers by the chi-square test. 4) Serum HBV DNA was detected in only one case out of 22 HBV carriers who cleared HBsAg during the follow-up period.in this case HBeAg was detected even after the appearance of anti-hbs.
468 羽 表1 全 症 例 の 血 中HBV関 田 元 連 マー カー と肝 組 織 中HBVDNAの 関係 * 非 癌 部 に つ い て も検 討 制限酵素未処理 右 端 の 数 字 はsize 図1 代 表 的 オ ー トラ ジオ グラ ム Hind markerを 示 す(単 位Kbp) IIIに 上 る 切 断 肝 細 胞 癌 に お け るHBVDNAの 組 込み HBV関 (症 例7)
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Key words: HBV carrier, anti-hd anti-hd IgM, HDAg HDVマ 1227 血 清 で陽 性)の2例 し た8). 疾 患 別 の 体 検 出 率 の 統 計 処 理 はx2検 成 体 検 出 率(Table 体 は,HBVキ (1.8%)に 検 出 さ れ た.疾 性 肝 疾 患62例 中6例 用:1例)感 体 は,慢 不 明 で あ る. ャ リ ア328例
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