Development and application of multiplex polymerase chain reaction for the etiological diagnosis of infectious endophthalmitis

• Main Authors: Bagyalakshmi R, Madhavan H, Therese K
• Format: Article
• Language: English
• Published: Wolters Kluwer Medknow Publications 2006-01-01
• Series: Journal of Postgraduate Medicine
• Subjects: Multiplex PCR; polymerase chain reaction; aqueous humor; vitreous fluid; infectious endophthalmitis
• Online Access: http://www.jpgmonline.com/article.asp?issn=0022-3859;year=2006;volume=52;issue=3;spage=179;epage=182;aulast=Bagyalakshmi

<b>Background: </b> Uniplex polymerase chain reaction (PCR) for detection of bacterial and panfungal genome has been applied onto a large number of intraocular fluids facilitating management of infective endophthalmitis. <b> Aim:</b> To develop and apply a novel, rapid multiplex polymerase chain reaction (mPCR) to detect the presence of eubacterial, <i> Propionibacterium acnes</i> and panfungal genomes in intraocular fluids from patients clinically diagnosed to have infective endophthalmitis. <b> Settings and Design: </b> Prospective study<b> . Materials and Methods:</b> Conventional methods of direct microscopy by KOH/calcofluor mount, Gram&#x2032;s staining and culture were done on 30 (19 Aqueous humor-AH and 11 Vitreous fluid-VF) intraocular specimens and mPCR done for simultaneous detection of eubacterial, <i> P. acnes</i> and panfungal genomes. <b> Results: </b> mPCR detected an infectious etiology in 18 (60&#x0025;) of 30 intraocular specimens. Eubacterial genome was detected in 12 (40&#x0025;) specimens, <i> P. acnes</i> genome in 4 (13.3&#x0025;) specimens and panfungal genome in 2 (6.6&#x0025;) specimens. mPCR results correlated with those of uniplex PCR. mPCR results were available within 5-6 hours after receipt of specimen, as against 8 hours required for each uniplex PCR with three separate thermalcyclers for their completion. Consumption of <i> Taq</i> polymerase was reduced considerably for mPCR. <b> Conclusion:</b> mPCR is a cost effective, single tube method for the simultaneous detection of eubacterial, <i> P. acnes</i> and panfungal genomes in intraocular specimens from patients with infective endophthalmitis. It is a more rapid procedure than uniplex PCRs and requires only a single thermalcycler.