Park Sunggyun, Kim Kyoungbo, Lee Youhyun, Ryoo Namhee
Departments of Laboratory Medicine, Keimyung University School of Medicine, Daegu 42601, Republic of Korea.
Departments of Ophthalmology, Keimyung University School of Medicine, Daegu 42601, Republic of Korea.
Diagnostics (Basel). 2025 Apr 10;15(8):966. doi: 10.3390/diagnostics15080966.
: Ocular infections can cause severe complications, including blindness, and distinguishing bacterial from fungal keratitis based on clinical features alone is difficult. This study compared broad-range conventional PCR and real-time PCR methods targeting the gene with traditional culture for diagnosing bacterial ocular infections. : We analyzed 160 ocular specimens from 111 patients, categorizing them as septic or aseptic. The results of both conventional PCR and real-time PCR methods targeting the gene were compared with traditional culture outcomes. : Real-time PCR demonstrated higher sensitivity than conventional PCR, and receiver operating characteristic analysis determined optimal ΔCT cutoff values of -2.13 and -4.09 for septic and aseptic specimens, respectively. Delays in specimen processing significantly affected real-time PCR accuracy. The meta-taxonomic analysis using nanopore sequencing only validated the PCR results when the DNA concentration was sufficient. : Broad-range real-time PCR proved to be a valuable diagnostic tool, particularly in aseptic specimens, with greater sensitivity and specificity than conventional PCR. The established ΔCT cutoff values improved diagnostic accuracy and showed that standardized specimen collection and processing are crucial for maximizing PCR efficacy.
眼部感染可导致包括失明在内的严重并发症，仅根据临床特征区分细菌性角膜炎和真菌性角膜炎很困难。本研究比较了针对该基因的广谱常规PCR和实时PCR方法与传统培养法在诊断细菌性眼部感染中的应用。我们分析了来自111例患者的160份眼部标本，将其分为感染性或非感染性。将针对该基因的常规PCR和实时PCR方法的结果与传统培养结果进行比较。实时PCR显示出比常规PCR更高的灵敏度，受试者工作特征分析确定感染性和非感染性标本的最佳ΔCT临界值分别为-2.13和-4.09。标本处理延迟显著影响实时PCR的准确性。仅当DNA浓度足够时，使用纳米孔测序的宏分类分析才验证了PCR结果。广谱实时PCR被证明是一种有价值的诊断工具，特别是在非感染性标本中，其灵敏度和特异性高于常规PCR。既定的ΔCT临界值提高了诊断准确性，并表明标准化的标本采集和处理对于最大化PCR功效至关重要。
