Chiquet Christophe, Lina Gérard, Benito Yvonne, Cornut Pierre-Loïc, Etienne Jérôme, Romanet Jean-Paul, Denis Philippe, Vandenesch François
Department of Ophthalmology, Hôpital Edouard Herriot, Université Lyon, France.
J Cataract Refract Surg. 2007 Apr;33(4):635-41. doi: 10.1016/j.jcrs.2006.12.017.
To identify bacterial agents in the aqueous humor of patients with postoperative endophthalmitis using eubacterial polymerase chain reaction (PCR) and conventional culture.
University Hospital of Lyon E. Herriot, Lyon, France.
Broad-range eubacterial PCR amplification followed by direct sequencing was used to identify microbial pathogens in ocular samples from 30 patients with acute or delayed-onset endophthalmitis, mainly after cataract surgery. Ocular samples included aqueous humor collected before the first intravitreal injection of antibiotics and vitreous samples collected at the time of the therapeutic pars plana vitrectomy.
Cultures were positive in 32% of cases and PCR in 61% of cases with aqueous humor samples. When associated, culture and PCR of aqueous humor samples allowed for a microbiological diagnosis in 71% of cases. Microorganisms cultured by conventional techniques matched those identified by PCR. When applied on vitreous pretreated with intravitreal antibiotics, PCR increased the identification rate from 18% to 62%.
Polymerase chain reaction assay of initial aqueous humor samples contributed to the diagnosis of endophthalmitis in 30% of cases. Previous use of intravitreal antibiotics did not seem to affect the ability to PCR-amplify DNA in the short term. Polymerase chain reaction-based technology was a useful adjunct to conventional culture because when used with aqueous humor samples only, the association of both techniques allowed for a microbiological diagnosis in 71% of cases of postoperative acute and delayed-onset endophthalmitis.
采用真细菌聚合酶链反应(PCR)和传统培养方法,鉴定术后眼内炎患者房水中的细菌病原体。
法国里昂埃里奥医院大学医院。
对30例急性或迟发性眼内炎患者(主要为白内障手术后)的眼部样本,采用广谱真细菌PCR扩增并直接测序来鉴定微生物病原体。眼部样本包括首次玻璃体腔内注射抗生素前采集的房水样本,以及治疗性玻璃体切割术时采集的玻璃体样本。
房水样本培养阳性率为32%,PCR阳性率为61%。房水样本培养和PCR联合应用时,71%的病例可作出微生物学诊断。传统技术培养的微生物与PCR鉴定的微生物相匹配。对经玻璃体腔内抗生素预处理的玻璃体样本进行检测时,PCR将鉴定率从18%提高到了62%。
初始房水样本的聚合酶链反应检测在30%的病例中有助于眼内炎的诊断。先前使用玻璃体腔内抗生素似乎短期内不影响PCR扩增DNA的能力。基于聚合酶链反应的技术是传统培养的有用辅助手段,因为仅与房水样本联合使用时,两种技术可使71%的术后急性和迟发性眼内炎病例作出微生物学诊断。