Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y, Masuda T
Department of Pathology, Iwate Medical University School of Medicine, Morioka 020-8505, Japan.
J Clin Pathol. 2003 Oct;56(10):778-9. doi: 10.1136/jcp.56.10.778.
To demonstrate the usefulness of polymerase chain reaction (PCR) methodology with both the FR2A/LJH/VLJH and the FR1c/LJH/VLJH primer sets for detecting monoclonal immunoglobulin heavy chain (IgH) gene rearrangement in B cell non-Hodgkin lymphomas (B-NHLs).
Eighty three patients with B-NHL were enrolled in this study. DNA was isolated from paraffin wax embedded sections and amplified by PCR to determine the sequences of the rearranged IgH gene. Each PCR product was subcloned. Cycle sequences and sequence analyses were done to determine the clone specific IgH variable region (VH) sequences.
Clonal IgH gene rearrangements were detected in 45 cases with FR2a/JH/VLJH and in 14 of the remaining cases with FR1c/JH/VLJH. Most of the cases detectable with FR2a/JH/VLJH were derived from VH3 and VH4 families. Five of six cases in the VH1 family and one in the VH7 family were amplified with the FR1c/JH/VLJH primer set only.
The detection rate of IgH rearrangement in B-NHLs can be increased by using both FR2A/LJH/VLJH and FR1c/LJH/VLJH, and these two primer sets are suitable for routine PCR methodology. Moreover, each primer set appears to be closely related to VH family specificity.
证明聚合酶链反应(PCR)方法使用FR2A/LJH/VLJH和FR1c/LJH/VLJH引物组检测B细胞非霍奇金淋巴瘤(B-NHL)中单克隆免疫球蛋白重链(IgH)基因重排的实用性。
83例B-NHL患者纳入本研究。从石蜡包埋切片中分离DNA,通过PCR扩增以确定重排IgH基因的序列。每个PCR产物进行亚克隆。进行循环测序和序列分析以确定克隆特异性IgH可变区(VH)序列。
使用FR2a/JH/VLJH在45例中检测到克隆性IgH基因重排,其余病例中使用FR1c/JH/VLJH在14例中检测到。大多数能用FR2a/JH/VLJH检测到的病例源自VH3和VH4家族。VH1家族的6例中有5例和VH7家族的1例仅用FR1c/JH/VLJH引物组扩增。
同时使用FR2A/LJH/VLJH和FR1c/LJH/VLJH可提高B-NHL中IgH重排的检测率,这两种引物组适用于常规PCR方法。此外,每种引物组似乎与VH家族特异性密切相关。