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Factors influencing polymerase chain reaction outcomes in patients with clinically suspected ocular tuberculosis

机译:临床怀疑眼结核患者中影响聚合酶链反应结果的因素

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Polymerase chain reaction (PCR) assay can be a useful method for definitive diagnosis in paucibacillary infections such as ocular tuberculosis (TB). In this study, we have evaluated factors affecting PCR outcomes in patients with clinically suspected ocular TB. Patients with clinically suspected ocular TB were investigated by PCR of aqueous or vitreous samples. Three control groups were also tested: group 1 included culture-proven non-tuberculous endophthalmitis, group 2 culture-negative non-tuberculous endophthalmitis, and group 3 patients undergoing surgery for uncomplicated cataract. PCR targeted one or more of following targets: IS6110, MPB64, and protein b genes of Mycobacterium tuberculosis complex. Multiple regression analysis (5% level of significance) was done to evaluate the associations between positive PCR outcome and laterality of disease, tuberculin skin test (TST)/interferon-gamma release assay (IGRA), chest radiography, and type of sample (aqueous or vitreous). The main outcome measures were positive PCR by one or more gene targets, and factors influencing positive PCR outcomes. All 114 samples were tested for MPB64, 110 for protein b, and 88 for IS6110. MPB64 was positive in 70.2% (n?=?80) of tested samples, protein b in 40.0% (n?=?44), and IS6110 in only 9.1% (n?=?8). DNA sequencing of amplicons from four randomly chosen PCR reactions showed homology for M. tuberculosis complex. Of the 80 PCR-positive patients, 71 completed a full course of antitubercular therapy, of which 65 patients (91.5%) had complete resolution of inflammation at final follow-up. Among controls, 12.5% (3 out of 24) in group 1 and 18.7% (6 out of 32) in group 2 also tested positive by PCR. No PCR-positive outcome was observed in control group 3 (n?=?25). Multiple regression analysis revealed significant association of positive PCR outcome with bilateral presentation, but not with a positive TST/IGRA, chest radiography, or type of sample (aqueous/vitreous) used. Careful selection of gene targets can yield high PCR positivity in clinically suspected ocular TB. Bilateral disease presentation but not any evidence of latent systemic TB influences PCR outcomes. False-positive results may be seen in ocular inflammation unrelated to ocular TB.
机译:聚合酶链反应(PCR)分析可以作为诊断诸如眼结核(TB)在内的脓疱感染的有用方法。在这项研究中,我们评估了影响临床可疑眼结核患者PCR结果的因素。临床上怀疑患有眼结核的患者通过PCR检测水样或玻璃样。还对三个对照组进行了测试:第1组包括经过培养证明的非结核性眼内炎;第2组是培养阴性的非结核性眼内炎;第3组因单纯性白内障而接受手术的患者。 PCR靶向以下一种或多种靶标:结核分枝杆菌复合物的IS6110,MPB64和蛋白b基因。进行了多元回归分析(显着性水平为5%),以评估PCR阳性结果与疾病的偏侧性,结核菌素皮肤试验(TST)/干扰素-γ释放试验(IGRA),胸部X线照片和样品类型(水性)之间的关联或玻璃体)。主要结果指标是通过一个或多个基因靶标进行PCR阳性反应,以及影响PCR阳性结果的因素。测试了所有114个样品的MPB64、110个蛋白b和88个IS6110。 MPB64在70.2%(n?=?80)的测试样品中为阳性,蛋白b在40.0%(n?=?44),而IS6110仅在9.1%(n?=?8)。从四个随机选择的PCR反应中扩增子的DNA测序表明结核分枝杆菌复合物具有同源性。在80位PCR阳性患者中,有71位完成了完整的抗结核治疗疗程,其中65位患者(91.5%)在最终随访中已完全消炎。在对照组中,第1组的12.5%(24个中的3个)和第2组的18.7%(32个中的6个)也通过PCR检测为阳性。在对照组3中未观察到PCR阳性结果(n≥25)。多元回归分析显示,PCR阳性结果与双侧表现显着相关,但与TST / IGRA阳性,胸部放射线照相或所用样品类型(水性/玻璃质)无关。仔细选择基因靶标可在临床怀疑的眼结核中产生高PCR阳性。双边疾病的表现,但没有任何潜在的系统性结核病的证据会影响PCR结果。在与眼结核无关的眼部炎症中可能会看到假阳性结果。

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