Sengler U, Reinhard T, Adams O, Gerlich W, Sundmacher R
Lions Cornea Bank North Rhine Westfalia, Eye Hospital, Heinrich-Heine University, Düsseldorf, Germany.
Graefes Arch Clin Exp Ophthalmol. 2001 Oct;239(10):783-7. doi: 10.1007/s004170100359.
The prevalence of donors seropositive for hepatitis B virus (HBV) surface antigen (HBsAg) or hepatitis C virus (HCV) antibody (anti-HCV) in western countries is estimated to be 0.5%-1%. There have been only two cases, however, published so far, where hepatitis B was suspected to have been transmitted by penetrating keratoplasty [4]. Concerning HCV, no suspected transmission by keratoplasty has been reported so far. This is also true for the time before serological screening for infectious diseases became mandatory for corneal donors. In the Lions Cornea Bank North Rhine Westfalia, 4.7% (HBV) and 3.2% (HCV) respectively of the corneas of the years 1995 to 1999 were discarded due to a "non-negative serology". In about 50% of these cases the screening test (ELISA) generated no valid signal and, therefore, a "questionable positivity" was assumed. Since in Germany corneal graft shortage still is a limiting factor for penetrating keratoplasty, this study was to evaluate the detectability of HBV-DNA and HCV-RNA in the serum samples, organ culture media and corneas of donors tested seropositive for HBsAg or anti-HCV in an attempt to obtain information as to the potential infectivity of this donor material. In this study, 29 corneas of 17 donors seropositive by ELISA for HBsAg and 27 corneas of 14 donors seropositive by ELISA for anti-HCV were evaluated. The organ culture media and the sera were screened for the presence of HBV-DNA or HCV-RNA by PCR. The corneoscleral discs were divided into a central trephinate (7 mm) and the corneoscleral rim. Concerning HBV-DNA both tissues were examined separately by polymerase chain reaction (PCR). In the case of HCV-RNA, a further, more sensitive nucleotide amplification method (NAT), the transcription mediated amplification (TMA), was used to test media, central corneas and cornealscleral rims. The media were additionally tested by PCR. Viral nucleic acid was detected in the sera from 6 of 17 HBsAg positive donors and from 6 of 14 anti-HCV positive donors. Viral genomes could not be detected in the organ culture media nor in the central corneas or corneoscleral rims by PCR at a detection limit of 1000 and 100 copies/ml. Concerning HCV-RNA, two media were positive in the TMA with 50-100 copies/ml.
according to our results, the risk of transmitting hepatitis B or C virus by penetrating keratoplasty appears to be low: although hepatitis C virus RNA could be detected in 2 media (from two donors) out of 27 with a low concentration of virus copies between 50-100/ml. It remains open whether such a low virus particle number may cause infection in the recipient.
据估计，西方国家乙肝病毒（HBV）表面抗原（HBsAg）或丙肝病毒（HCV）抗体（抗-HCV）血清学阳性的供体患病率为0.5%-1%。然而，迄今为止仅发表过两例疑似穿透性角膜移植传播乙肝的病例[4]。关于丙肝，目前尚无角膜移植疑似传播的报道。在传染病血清学筛查成为角膜供体强制要求之前也是如此。在北莱茵-威斯特法伦州狮子角膜库，1995年至1999年期间分别有4.7%（乙肝）和3.2%（丙肝）的角膜因“血清学非阴性”而被废弃。在这些病例中，约50%的筛查试验（酶联免疫吸附测定法，ELISA）未产生有效信号，因此被认为是“可疑阳性”。由于在德国，角膜移植材料短缺仍是穿透性角膜移植的限制因素，本研究旨在评估在ELISA检测中HBsAg或抗-HCV血清学阳性供体的血清样本、器官培养基和角膜中HBV-DNA和HCV-RNA的可检测性，以获取有关这种供体材料潜在传染性的信息。在本研究中，对17例ELISA检测HBsAg阳性供体的29只角膜和14例ELISA检测抗-HCV阳性供体的27只角膜进行了评估。通过聚合酶链反应（PCR）筛查器官培养基和血清中是否存在HBV-DNA或HCV-RNA。将角膜巩膜盘分为中央环钻组织（7毫米）和角膜巩膜边缘。对于HBV-DNA，两种组织分别通过聚合酶链反应（PCR）进行检测。对于HCV-RNA，采用另一种更灵敏的核苷酸扩增方法——转录介导扩增（TMA）来检测培养基、中央角膜和角膜巩膜边缘。培养基还通过PCR进行检测。在17例HBsAg阳性供体的6例血清以及14例抗-HCV阳性供体的6例血清中检测到病毒核酸。在检测限为1000和100拷贝/毫升时，通过PCR在器官培养基、中央角膜或角膜巩膜边缘均未检测到病毒基因组。关于HCV-RNA，在TMA检测中有两种培养基呈阳性，病毒拷贝数浓度为50-100拷贝/毫升。
根据我们的结果，穿透性角膜移植传播乙肝或丙肝病毒的风险似乎较低：尽管在27份样本中的2份培养基（来自两名供体）中检测到HCV-RNA，病毒拷贝数浓度较低，为50-100/毫升。如此低的病毒颗粒数是否会导致受者感染仍不明确。
