The diagnosis of delayed post-operative endophthalmitis by polymerase chain reaction of bacterial DNA in vitreous samples

1 Departments of Clinical Science, Institute of Ophthalmology, Bath Street, London EC1V 9EL * Department of Medical Microbiology. University College London Medical School, London W1P 7PN Departments of Pathology, Institute of Ophthalmology, Bath Street, London EC1V 9EL Received October 1, 1993 Accep...

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Veröffentlicht in:Journal of medical microbiology 1994-06, Vol.40 (6), p.408-415
Hauptverfasser: Hykin, P. G, Tobal, K, McIntyre, G, Matheson, M. M, Towler, H. M. A, Lightman, S. L
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container_end_page 415
container_issue 6
container_start_page 408
container_title Journal of medical microbiology
container_volume 40
creator Hykin, P. G
Tobal, K
McIntyre, G
Matheson, M. M
Towler, H. M. A
Lightman, S. L
description 1 Departments of Clinical Science, Institute of Ophthalmology, Bath Street, London EC1V 9EL * Department of Medical Microbiology. University College London Medical School, London W1P 7PN Departments of Pathology, Institute of Ophthalmology, Bath Street, London EC1V 9EL Received October 1, 1993 Accepted December 9, 1993 Delayed post-operative endophthalmitis is a complication of modern cataract extraction and posterior chamber lens implantation. Propionibacterium acnes has been isolated in a few such cases but the majority are culture-negative, compounding surgical and medical management decisions. A method of detecting bacterial, and specifically P. acnes , DNA by the polymerase chain reaction (PCR) directed at 16S rDNA is reported. Nested PCR with universal eubacterial primers complimentary to regions of 16S rDNA conserved sequences detected 50 fg of bacterial DNA spike in normal vitreous. Nested PCR with P. acnes primers detected 10 fg of DNA. Vitreous samples from 29 patients undergoing vitrectomy for reasons unrelated to infection and 23 samples from 19 patients with delayed post-operative endophthalmitis were analysed. Four (14%) of 29 normal individuals and 17 (74%) of 23 delayed cases gave positive results with universal eubacterial primers. None of 29 and eight of 23 samples gave positive results with P. acnes primers. The 14% positive rate with universal primers in non-infected cases may limit their use in delayed post-operative endophthalmitis. PCR detection of bacterial DNA with specific primers from vitreous samples may prove a useful means of diagnosing delayed post-operative endophthalmitis and facilitating management decisions when conventional bacterial culture is negative.
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Nested PCR with universal eubacterial primers complimentary to regions of 16S rDNA conserved sequences detected 50 fg of bacterial DNA spike in normal vitreous. Nested PCR with P. acnes primers detected 10 fg of DNA. Vitreous samples from 29 patients undergoing vitrectomy for reasons unrelated to infection and 23 samples from 19 patients with delayed post-operative endophthalmitis were analysed. Four (14%) of 29 normal individuals and 17 (74%) of 23 delayed cases gave positive results with universal eubacterial primers. None of 29 and eight of 23 samples gave positive results with P. acnes primers. The 14% positive rate with universal primers in non-infected cases may limit their use in delayed post-operative endophthalmitis. 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G</creatorcontrib><creatorcontrib>Tobal, K</creatorcontrib><creatorcontrib>McIntyre, G</creatorcontrib><creatorcontrib>Matheson, M. M</creatorcontrib><creatorcontrib>Towler, H. M. A</creatorcontrib><creatorcontrib>Lightman, S. L</creatorcontrib><title>The diagnosis of delayed post-operative endophthalmitis by polymerase chain reaction of bacterial DNA in vitreous samples</title><title>Journal of medical microbiology</title><addtitle>J Med Microbiol</addtitle><description>1 Departments of Clinical Science, Institute of Ophthalmology, Bath Street, London EC1V 9EL * Department of Medical Microbiology. University College London Medical School, London W1P 7PN Departments of Pathology, Institute of Ophthalmology, Bath Street, London EC1V 9EL Received October 1, 1993 Accepted December 9, 1993 Delayed post-operative endophthalmitis is a complication of modern cataract extraction and posterior chamber lens implantation. Propionibacterium acnes has been isolated in a few such cases but the majority are culture-negative, compounding surgical and medical management decisions. A method of detecting bacterial, and specifically P. acnes , DNA by the polymerase chain reaction (PCR) directed at 16S rDNA is reported. Nested PCR with universal eubacterial primers complimentary to regions of 16S rDNA conserved sequences detected 50 fg of bacterial DNA spike in normal vitreous. Nested PCR with P. acnes primers detected 10 fg of DNA. Vitreous samples from 29 patients undergoing vitrectomy for reasons unrelated to infection and 23 samples from 19 patients with delayed post-operative endophthalmitis were analysed. Four (14%) of 29 normal individuals and 17 (74%) of 23 delayed cases gave positive results with universal eubacterial primers. None of 29 and eight of 23 samples gave positive results with P. acnes primers. The 14% positive rate with universal primers in non-infected cases may limit their use in delayed post-operative endophthalmitis. PCR detection of bacterial DNA with specific primers from vitreous samples may prove a useful means of diagnosing delayed post-operative endophthalmitis and facilitating management decisions when conventional bacterial culture is negative.</description><subject>Bacterial diseases</subject><subject>Bacterial diseases of the eye and orbit</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Conserved Sequence</subject><subject>DNA Primers - chemistry</subject><subject>DNA, Bacterial - analysis</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA, Ribosomal - analysis</subject><subject>DNA, Ribosomal - chemistry</subject><subject>Endophthalmitis - diagnosis</subject><subject>Eye Infections, Bacterial - diagnosis</subject><subject>Gram-Positive Bacterial Infections - diagnosis</subject><subject>Human bacterial diseases</subject><subject>Humans</subject><subject>Infectious diseases</subject><subject>Medical sciences</subject><subject>Molecular Sequence Data</subject><subject>Polymerase Chain Reaction</subject><subject>Postoperative Complications - diagnosis</subject><subject>Propionibacterium acnes - genetics</subject><subject>Propionibacterium acnes - isolation &amp; purification</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>Sensitivity and Specificity</subject><subject>Vitrectomy</subject><subject>Vitreous Body - microbiology</subject><issn>0022-2615</issn><issn>1473-5644</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kM1u3CAUhVHVKp2mfYAuKrGo2pVTMGCbZZT0T4qaTbpGYC5jImNc8CTy2wdrprM5IJ3vnnt1EPpIyRUlUn4jpK7rhoqKk6op0r1CO8pbVomG89dot_nVBrxF73J-JIS2jMkLdNER0kjGdmh9GABbr_dTzD7j6LCFUa9g8RzzUsUZkl78E2CYbJyHZdBj8EshzVqIcQ3Fz4D7QfsJJ9D94uO0xZjyheT1iG__XONiPvklQTxknHWYR8jv0RunxwwfTu8l-vvj-8PNr-ru_ufvm-u7qmdUdBU1tG-ZkWDahhlrwNbGQUuckLQlnNFWWls7V7QFYgV3upZMaiFoBwwadom-HHPnFP8dIC8q-NzDOOppO0e1jRBbSgHpEexTzDmBU3PyQadVUaK2utX_uhUnqinSlZlPp_CDCWDPE6d-i__55Ovc69ElPfU-nzFOGadcFOzrERv8fnj2CdQepuDLIcZH9RjCeeELnCGXVA</recordid><startdate>199406</startdate><enddate>199406</enddate><creator>Hykin, P. 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Propionibacterium acnes has been isolated in a few such cases but the majority are culture-negative, compounding surgical and medical management decisions. A method of detecting bacterial, and specifically P. acnes , DNA by the polymerase chain reaction (PCR) directed at 16S rDNA is reported. Nested PCR with universal eubacterial primers complimentary to regions of 16S rDNA conserved sequences detected 50 fg of bacterial DNA spike in normal vitreous. Nested PCR with P. acnes primers detected 10 fg of DNA. Vitreous samples from 29 patients undergoing vitrectomy for reasons unrelated to infection and 23 samples from 19 patients with delayed post-operative endophthalmitis were analysed. Four (14%) of 29 normal individuals and 17 (74%) of 23 delayed cases gave positive results with universal eubacterial primers. None of 29 and eight of 23 samples gave positive results with P. acnes primers. The 14% positive rate with universal primers in non-infected cases may limit their use in delayed post-operative endophthalmitis. PCR detection of bacterial DNA with specific primers from vitreous samples may prove a useful means of diagnosing delayed post-operative endophthalmitis and facilitating management decisions when conventional bacterial culture is negative.</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>8006933</pmid><doi>10.1099/00222615-40-6-408</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
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subjects Bacterial diseases
Bacterial diseases of the eye and orbit
Base Sequence
Biological and medical sciences
Conserved Sequence
DNA Primers - chemistry
DNA, Bacterial - analysis
DNA, Bacterial - chemistry
DNA, Ribosomal - analysis
DNA, Ribosomal - chemistry
Endophthalmitis - diagnosis
Eye Infections, Bacterial - diagnosis
Gram-Positive Bacterial Infections - diagnosis
Human bacterial diseases
Humans
Infectious diseases
Medical sciences
Molecular Sequence Data
Polymerase Chain Reaction
Postoperative Complications - diagnosis
Propionibacterium acnes - genetics
Propionibacterium acnes - isolation & purification
RNA, Ribosomal, 16S - genetics
Sensitivity and Specificity
Vitrectomy
Vitreous Body - microbiology
title The diagnosis of delayed post-operative endophthalmitis by polymerase chain reaction of bacterial DNA in vitreous samples
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