Falk K, Gratama J W, Rowe M, Zou J Z, Khanim F, Young L S, Oosterveer M A, Ernberg I
Microbiology and Tumorbiology Center, Karolinska Institute, Stockholm, Sweden.
J Gen Virol. 1995 Apr;76 ( Pt 4):779-90. doi: 10.1099/0022-1317-76-4-779.
The six Epstein-Barr virus (EBV) nuclear antigen proteins (EBNA-1-6) show characteristic size variations between different virus isolates; this is a feature that has been used to identify the source of virus isolates in epidemiological studies (Ebnotyping). We have now studied the correlation between restriction fragment length polymorphisms (RFLPs) within exons coding for the EBNAs and the molecular masses of the respective proteins. The B95-8 EBV strain was used as the prototype virus. The variation in apparent molecular mass of EBNA-1, -3 and -6 correlated positively with the size of RFLP coding for repeat sequences in these polypeptides. For EBNA-2, no correlation between apparent molecular mass and length of the repetitive sequences was found. The EBNA-4 protein showed virtually no variation in apparent molecular mass and RFLP size across the repeat sequence. Based on the strong correlation between apparent molecular mass and RFLP size for EBNA-6, we developed an EBNA-6 PCR assay that discriminated between different isolates of EBV. This assay offers the advantage of EBV characterization using uncultured material (e.g. throat washings, blood or biopsies), thus avoiding the selection against poorly transforming strains that occurs during establishment of lymphoblastoid cell lines required for Ebnotyping at the protein level.
六种爱泼斯坦-巴尔病毒（EBV）核抗原蛋白（EBNA-1 - 6）在不同病毒分离株之间呈现出特征性的大小差异；这一特征已被用于在流行病学研究中鉴定病毒分离株的来源（EB病毒分型）。我们现在研究了编码EBNA的外显子内的限制性片段长度多态性（RFLP）与各蛋白分子量之间的相关性。B95 - 8 EBV株被用作原型病毒。EBNA-1、-3和-6的表观分子量变化与编码这些多肽中重复序列的RFLP大小呈正相关。对于EBNA-2，未发现表观分子量与重复序列长度之间存在相关性。EBNA-4蛋白在整个重复序列中的表观分子量和RFLP大小几乎没有变化。基于EBNA-6的表观分子量与RFLP大小之间的强相关性，我们开发了一种EBNA-6 PCR检测方法，可区分不同的EBV分离株。该检测方法的优点是可使用未培养的材料（如咽洗液、血液或活检组织）对EBV进行特征分析，从而避免了在蛋白质水平进行EB病毒分型所需的淋巴母细胞系建立过程中对转化能力差的菌株的选择。
