Sharma Surya Prakash, Sharma Ashok, Dwivedi Sujata, Sharma Anil, Sharma Ravi Kant, Datta Sonal
Advanced Eye Centre, Department of Ophthalmology, Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India.
Department of Bio-Sciences and Technology, MMEC, Maharishi Markandeshwar (Deemed to be University), Mullana-Ambala, Haryana, India.
Int Ophthalmol. 2025 Jul 21;45(1):300. doi: 10.1007/s10792-025-03609-z.
Infective keratitis is a serious ocular condition where early and accurate identification of the causative organisms is crucial for effective treatment. However, traditional diagnostic methods such as microbial culture often have low sensitivity, particularly in microbial culture-negative cases.
This study aimed to assess and compare the performance of conventional methods (microbial culture and smear microscopy) with molecular methods (PCR) for detecting bacterial and fungal pathogens in patients with infective keratitis in North India.
A total of 109 patients with clinically diagnosed infective keratitis and 30 healthy controls were enrolled between January 2023 and January 2024. Diagnostic methods included slit-lamp examination, microbial culture, 10% KOH wet mount, Gram staining, and polymerase chain reaction (PCR). The sensitivity and specificity of each method were evaluated.
Microbial culture showed a sensitivity of 14.68% and specificity of 90%, with Aspergillus and Staphylococcus species being the most commonly isolated organisms. Smear microscopy showed better sensitivity (72.47%). PCR achieved the highest sensitivity (84.4%) and detected pathogens in 86 out of 120 culture-negative cases. PCR identified 57 fungal and 48 bacterial infections, including 10 cases of mixed infections. 10% KOH smear detected fungi in 55 samples, and Gram stain identified bacteria in 23 samples.
PCR significantly improved pathogen detection, especially in culture-negative and polymicrobial cases. The findings support the integration of molecular diagnostics with conventional methods for more accurate and timely diagnosis of infective keratitis.
感染性角膜炎是一种严重的眼部疾病,早期准确识别致病微生物对于有效治疗至关重要。然而,传统的诊断方法如微生物培养往往敏感性较低,尤其是在微生物培养阴性的病例中。
本研究旨在评估和比较传统方法(微生物培养和涂片显微镜检查)与分子方法(PCR)在检测印度北部感染性角膜炎患者细菌和真菌病原体方面的性能。
2023年1月至2024年1月期间,共纳入109例临床诊断为感染性角膜炎的患者和30例健康对照。诊断方法包括裂隙灯检查、微生物培养、10%氢氧化钾湿片法、革兰氏染色和聚合酶链反应(PCR)。评估了每种方法的敏感性和特异性。
微生物培养的敏感性为14.68%,特异性为90%,最常分离出的微生物是曲霉菌和葡萄球菌。涂片显微镜检查显示出更好的敏感性(72.47%)。PCR的敏感性最高(84.4%),在120例培养阴性的病例中检测到86例病原体。PCR鉴定出57例真菌感染和48例细菌感染,包括10例混合感染。10%氢氧化钾涂片在55个样本中检测到真菌,革兰氏染色在23个样本中鉴定出细菌。
PCR显著提高了病原体检测率,尤其是在培养阴性和多微生物感染的病例中。这些发现支持将分子诊断与传统方法相结合,以更准确、及时地诊断感染性角膜炎。