Somerville Tobi F, Mdala Shaffi, Zungu Thokozani, Gandiwa Moira, Herbert Rose, Everett Dean, Corless Caroline E, Beare Nicholas A V, Neal Timothy, Horsburgh Malcolm J, Darby Alistair, Kaye Stephen B, Kayange Petros C
Department of Eye and Vision Sciences, University of Liverpool, Liverpool, UK
Queen Elizabeth Central Hospital, Blantyre, Southern Region, Malawi.
BMJ Open Ophthalmol. 2024 Apr 22;9(1):e001682. doi: 10.1136/bmjophth-2024-001682.
Microbial keratitis (MK) is a significant cause of blindness in sub-Saharan Africa. We investigated the feasibility of using a novel corneal impression membrane (CIM) for obtaining and processing samples by culture, PCR and whole-genome sequencing (WGS) in patients presenting with suspected MK in Malawi.
Samples were collected from patients presenting with suspected MK using a 12 mm diameter polytetrafluoroethylene CIM disc. Samples were processed using culture and PCR for , herpes simplex virus type 1 (HSV-1) and the bacterial 16S rRNA gene. Minimum inhibitory concentrations of isolates to eight antimicrobials were measured using susceptibility strips. WGS was used to characterise isolates.
71 eyes of 71 patients were included. The overall CIM isolation rate was 81.7% (58 positive samples from 71 participants). 69 (81.2%) of isolates were Gram-positive cocci. Coagulase-negative 31.8% and species 14.1% were the most isolated bacteria. Seven (9.9%) participants were positive for HSV-1. Fungi and were not detected. Moxifloxacin and chloramphenicol offered the best coverage for both Gram-positive and Gram-negative isolates when susceptibility was determined using known antimicrobial first quartile concentrations and European Committee on Antimicrobial Susceptibility Testing breakpoints, respectively. WGS identified known virulence genes associated with keratitis.
In a resource-poor setting, a CIM can be used to safely sample the cornea in patients presenting with suspected MK, enabling identification of causative microorganisms by culture and PCR. Although the microbiological spectrum found was limited to the dry season, these preliminary results could be used to guide empirical treatment.
微生物性角膜炎(MK)是撒哈拉以南非洲地区失明的一个重要原因。我们调查了在马拉维疑似患有MK的患者中,使用新型角膜印模膜(CIM)通过培养、聚合酶链反应(PCR)和全基因组测序(WGS)获取和处理样本的可行性。
使用直径12毫米的聚四氟乙烯CIM盘从疑似患有MK的患者中采集样本。样本通过培养和PCR检测1型单纯疱疹病毒(HSV-1)和细菌16S核糖体RNA基因进行处理。使用药敏试纸测量分离株对8种抗菌药物的最低抑菌浓度。WGS用于鉴定分离株。
纳入了71例患者的71只眼。CIM的总体分离率为81.7%(71名参与者中有58个阳性样本)。69株(81.2%)分离株为革兰氏阳性球菌。凝固酶阴性葡萄球菌占31.8%,金黄色葡萄球菌占14.1%,是最常分离出的细菌。7名(9.9%)参与者HSV-1检测呈阳性。未检测到真菌和分枝杆菌。当分别使用已知抗菌药物的第一四分位数浓度和欧洲抗菌药物敏感性测试委员会的断点确定药敏性时,莫西沙星和氯霉素对革兰氏阳性和革兰氏阴性分离株的覆盖效果最佳。WGS鉴定出了与MK相关的已知毒力基因。
在资源匮乏的环境中,CIM可用于对疑似患有MK的患者的角膜进行安全采样,通过培养和PCR鉴定致病微生物。尽管发现的微生物谱仅限于旱季,但这些初步结果可用于指导经验性治疗。