Sugita Sunao, Takase Hiroshi, Sugamoto Yoshiharu, Arai Ayako, Miura Osamu, Mochizuki Manabu
Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan.
Jpn J Ophthalmol. 2009 May;53(3):209-14. doi: 10.1007/s10384-009-0662-y. Epub 2009 May 31.
To determine whether a diagnosis of intraocular lymphoma (IOL) can be made using a combination of polymerase chain reaction (PCR) analysis to detect gene rearrangement of immunoglobulin and cytokine concentrations in the vitreous fluid.
Vitreous samples from 22 patients with clinically suspected IOL and ten control patients with acute retinal necrosis or cytomegalovirus retinitis were examined by PCR analysis and cytokine measurements. Genomic DNA was extracted from the cells in the vitreous, and the immunoglobulin heavy chain (IgH) gene was amplified by two PCR procedures: (1) microdissection and PCR to detect IgH gene rearrangement and (2) qualitative PCR to detect IgH VDJ gene rearrangement. The supernatants of the vitreous samples were used for enzyme-linked immunosorbent assay to determine interleukin (IL)-10 and IL-6 levels.
PCR examinations detected IgH rearrangement in the vitreous in 21 of the 22 IOL patients (95.5%) and in none of the ten control patients. Elevated IL-10 concentrations (>100 pg/ml) and the IL-10/IL-6 ratio (>1.0) were positive in 18 of the 22 IOL patients (81.8%), but negative in all of the control patients. Sensitivity, specificity, positive predictive value, and negative predictive value of PCR for the diagnosis of IOL were calculated to be 0.955, 1.000, 1.000, and 0.909, respectively, and those of the cytokine concentration assay to be 0.818, 1.000, 1.000, and 0.714, respectively. When both the intravitreal cytokine assay and PCR analysis of the vitreous samples are used, as well as diagnostic criteria of IOL defined as a positive outcome from one of the two assays together with clinical signs, the sensitivity and specificity of the criteria were 1.000.
A combination of PCR assay to detect gene rearrangement of IgH and cytokine profiling (IL-10 and IL-6) is extremely useful for the diagnosis of intraocular lymphoma.
确定是否可通过聚合酶链反应（PCR）分析检测玻璃体中免疫球蛋白基因重排以及细胞因子浓度相结合的方法来诊断眼内淋巴瘤（IOL）。
对22例临床疑似IOL患者和10例急性视网膜坏死或巨细胞病毒性视网膜炎对照患者的玻璃体样本进行PCR分析和细胞因子检测。从玻璃体中的细胞提取基因组DNA，通过两种PCR方法扩增免疫球蛋白重链（IgH）基因：（1）显微切割和PCR以检测IgH基因重排；（2）定性PCR以检测IgH VDJ基因重排。玻璃体样本的上清液用于酶联免疫吸附测定以确定白细胞介素（IL）-10和IL-6水平。
PCR检测发现22例IOL患者中有21例（95.5%）玻璃体存在IgH重排，而10例对照患者均未检测到。22例IOL患者中有18例（81.8%）IL-10浓度升高（>100 pg/ml）且IL-10/IL-6比值>1.0，而所有对照患者均为阴性。PCR诊断IOL的敏感性、特异性、阳性预测值和阴性预测值分别计算为0.955、1.000、1.000和0.909，细胞因子浓度检测的相应值分别为0.818、1.000、1.000和0.714。当同时使用玻璃体内细胞因子检测和玻璃体样本的PCR分析，以及将两种检测之一呈阳性结果并伴有临床体征定义为IOL的诊断标准时，该标准的敏感性和特异性均为1.000。
检测IgH基因重排的PCR检测与细胞因子分析（IL-10和IL-6）相结合对眼内淋巴瘤的诊断极为有用。
