Application of in situ hybridization technique for quantitative assessment of ongoing symptomatic Epstein-Barr virus infection after living related liver transplantation.
Application of in situ hybridization technique for quantitative assessment of ongoing symptomatic Epstein-Barr virus infection after living related liver transplantation.
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应用原位杂交技术定量评估活体肝移植后持续症状性 Epstein-Barr 病毒感染。
DOI：
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发表时间：
1998
影响因子：
2.1
通讯作者：
Koichi Tanaka
中科院分区：
文献类型：
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作者：
Hiroto Egawa;Tsutomu Oh;Takashi Arai;Y. Inomata;Shinji Uemoto;K. Asonuma;Tetsuya Kinchi;Hideaki Okajima;Akira Matsui;Naomi Kawashima;Olivia M. Martinez;Koichi Tanaka
为了定量评估儿童肝移植受者持续的症状性eb病毒（EBV）感染，我们采用原位杂交（ISH）方法测定了感染EBV的外周血单个核细胞（PBMC）的数量，并将结果与这些患者的临床病程联系起来。1995年2月至1996年3月间，24例患者出现EBV感染症状。24例患者急性期采血，13例患者恢复期采血，37例患儿移植前采血。对PBMC进行ISH，对全血DNA进行聚合酶链反应（PCR）。从ebv编码的小核RNA 1 （EBER1）的编码序列中选择ISH寡核苷酸探针。在许多表达EBER1/5 × 104 PBMC （#EBER1）的细胞中报道了ISH的结果。发热、腹泻、上呼吸道症状、胸腔积液、腹水、淋巴结病变、淋巴增殖性疾病（LPD）伴有EBV感染，经血清学、病毒特异性染色或PCR证实为EBV相关疾病（EBVD）。所有EBER1阳性的样本均伴有EBV PCR阳性。#EBERI在急性期为68.2 +/- 144.9（平均+/- SD），范围为0 ~ 621，恢复期为0.20 +/- 0.41，范围为0 ~ 2，术前血清阳性23例患者为0.27 +/- 0.77，术前血清阴性14例患者均为0。EBVD患者的#EBER1显著高于恢复期和移植前患者。#EBERI大于10的患者的康复机会明显低于#EBERI小于10的患者，死亡率更高。我们得出结论，#EBER1可能是EBVD的特异性和定量标记物，并可能预测进展为LPD。
For quantitative assessment of ongoing symptomatic Epstein-Barr virus (EBV) infection in pediatric recipients of liver transplantation, we determined the number of peripheral blood mononuclear cells (PBMC) infected by EBV by in situ hybridization (ISH) and related the results with clinical courses of those patients. Twenty-four patients had symptomatic EBV infection between February 1995 and March 1996. Blood samples were obtained from these 24 patients at the time of acute phase, from 13 of them during convalescence, and 37 pediatric patients before transplantation. ISH was performed on the PBMC and polymerase chain reaction (PCR) on DNA from whole blood. Oligonucleotide probes for ISH were chosen from coding sequences of EBV-encoded small nuclear RNA 1 (EBER1). Results of ISH were reported in a number of cells expressing EBER1/5 x 104 PBMC (#EBER1). Fever, diarrhea, upper respiratory symptoms, pleural effusion, ascites, lymphadenopathy, and lymphoproliferative disease (LPD) accompanied with EBV infection proven by serology, viral-specific stain or PCR were regarded as EBV related diseases (EBVD). All samples with positive #EBER1 were accompanied by positive EBV PCR. #EBERI was 68.2 +/- 144.9 (mean +/- SD) ranging from 0 to 621 in the acute phase, 0.20 +/- 0.41 ranging from 0 to 2 in the convalescence phase, 0.27 +/- 0.77 in 23 preoperative patients with positive serology, and 0 in all 14 preoperative patients with negative serology. The #EBER1 in ongoing EBVD was significantly greater than that of patients in convalescence or before transplantation. Patients with #EBERI greater than 10 had a significantly lower chance of convalescence and a higher mortality than patients with #EBER 1 less than 10. We conclude that #EBER1 could be a specific and quantitative marker of EBVD and might predict progression to LPD.