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聚合酶链反应在细菌性眼内炎诊断中的应用

Polymerase chain reaction in the diagnosis of bacterial endophthalmitis.

作者信息

Therese K L, Anand A R, Madhavan H N

机构信息

Vision Research Foundation, Chennai, India.

出版信息

Br J Ophthalmol. 1998 Sep;82(9):1078-82. doi: 10.1136/bjo.82.9.1078.

Abstract

BACKGROUND

Microbiological investigations of vitreous fluid (VF) and aqueous humour (AH) specimens have often failed to detect the infecting agent in infectious endophthalmitis, resulting in a clinical dilemma regarding therapy. In this study, the polymerase chain reaction (PCR) was evaluated in the diagnosis of bacterial and Propionibacterium acnes endophthalmitis.

METHODS

58 intraocular specimens (30 VF and 28 AH) from 55 cases of endophthalmitis and 20 specimens (14 VF and 6 AH) as controls from non-infective disorders were processed for microbiological investigations. Nested PCR directed at the 16S rDNA using universal primers for eubacterial genome was done. PCR for P acnes was performed on specimens microbiologically negative by conventional techniques but eubacterial genome positive.

RESULTS

Of the 20 controls from non-infective cases, one (5%) was positive using eubacterial primers and none with P acnes primers. PCR for eubacterial genome showed 100% correlation with 20 (34.5%) bacteriologically positive specimens. Eubacterial genome, was detected in 17 (44.7%) of 38 bacteriologically negative specimens and nine (52.9%) out of the 17 were positive for P acnes genome. Among the 21 eubacterial PCR negative specimens, seven were fungus positive. By inclusion of PCR, microbiologically positive specimens increased from 46.5% to 75.8%. PCR on AH was as sensitive as that on VF for the detection of both eubacterial and the P acnes genome.

CONCLUSION

PCR performed on AH and VF is a reliable tool for the diagnosis of bacterial and P acnes endophthalmitis particularly in smear and culture negative specimens.

摘要

背景

对玻璃体液(VF)和房水(AH)标本进行微生物学检查时,常常无法在感染性眼内炎中检测到感染病原体,这导致了治疗方面的临床困境。在本研究中,对聚合酶链反应(PCR)在细菌性和痤疮丙酸杆菌性眼内炎诊断中的应用进行了评估。

方法

对来自55例眼内炎患者的58份眼内标本(30份VF和28份AH)以及作为对照的20份来自非感染性疾病的标本(14份VF和6份AH)进行微生物学检查。使用针对真细菌基因组的通用引物进行针对16S rDNA的巢式PCR。对常规技术微生物学检查为阴性但真细菌基因组为阳性的标本进行痤疮丙酸杆菌的PCR检测。

结果

在20份来自非感染性病例的对照标本中,使用真细菌引物检测到1份(5%)呈阳性,使用痤疮丙酸杆菌引物检测无一例呈阳性。真细菌基因组的PCR检测与20份(34.5%)细菌学检查呈阳性的标本具有100%的相关性。在38份细菌学检查为阴性的标本中,检测到17份(44.7%)真细菌基因组呈阳性,其中9份(52.9%)痤疮丙酸杆菌基因组呈阳性。在21份真细菌PCR检测为阴性的标本中,7份真菌检测呈阳性。通过纳入PCR检测,微生物学检查呈阳性的标本从46.5%增加到75.8%。AH上的PCR检测真细菌和痤疮丙酸杆菌基因组的敏感性与VF上的检测相同。

结论

对AH和VF进行PCR检测是诊断细菌性和痤疮丙酸杆菌性眼内炎的可靠工具,尤其是在涂片和培养均为阴性的标本中。

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