Ocular pathogen or commensal: A PCR-based study of surface bacterial flora in normal and dry eyes

Joanna E. Graham, Johnny Moore, Xu Jiru, John E. Moore, Edward A. Goodall, James S. G. Dooley, Velma Hayes, Darlene A. Dartt, Stephen Downes, Tara Moore

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

PURPOSE. To compare the bacterial population of the ocular surface of normal and dry eye subjects using conventional culture and 16S rDNA PCR. METHODS. Ninety-one subjects were classified as normal (n = 57) or dry eye (n = 34) by using tear break-up time, McMonnies survey, goblet cell density, and meibomian gland assessment. Conventional bacterial culture and broad-range 16S rDNA PCR, cloning, and DNA sequencing were used for bacterial identification. Repeated sampling was performed in a subset of subjects over a 3-month period. The association between goblet cell loss and bacterial counts in a subgroup of subjects was assessed. RESULTS. Most of the bacteria identified by culture were coagulase negative staphylococci, whereas molecular methods demonstrated a considerable number of additional bacteria. Atypical ocular surface bacteria including Rhodococcus erythropolis, Klebsiella oxytoca, and Erwinia sp., were identified in cases of overt inflammation and, surprisingly, on the normal ocular surface. The same bacteria remained on the ocular surface after repeated sampling. Increased bacterial flora was associated with reduced goblet cell density. CONCLUSIONS. Molecular analysis revealed a diverse ocular surface bacterial population. In addition to the normal flora, various potentially pathogenic bacteria were identified. The detection of known pathogens in both normal and dry eyes, with minimal signs of infection, presents a diagnostic dilemma. It remains unknown whether their presence is associated with inflammation and reduced goblet cell density or whether they adversely affect the ocular surface predisposing it to abnormal microbial colonization. In the absence of overt clinical infection, it is unknown whether such results should prompt intervention with therapy.
LanguageEnglish
Pages5616-5623
JournalINVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
Volume48
Issue number12
DOIs
Publication statusPublished - 1 Dec 2007

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Polymerase Chain Reaction
Goblet Cells
Bacteria
Cell Count
Ribosomal DNA
Meibomian Glands
Klebsiella oxytoca
Erwinia
Inflammation
Rhodococcus
Bacterial Load
Coagulase
Infection
Tears
Staphylococcus
DNA Sequence Analysis
Population
Organism Cloning

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Graham, Joanna E. ; Moore, Johnny ; Jiru, Xu ; Moore, John E. ; Goodall, Edward A. ; Dooley, James S. G. ; Hayes, Velma ; Dartt, Darlene A. ; Downes, Stephen ; Moore, Tara. / Ocular pathogen or commensal: A PCR-based study of surface bacterial flora in normal and dry eyes. In: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE. 2007 ; Vol. 48, No. 12. pp. 5616-5623.
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abstract = "PURPOSE. To compare the bacterial population of the ocular surface of normal and dry eye subjects using conventional culture and 16S rDNA PCR. METHODS. Ninety-one subjects were classified as normal (n = 57) or dry eye (n = 34) by using tear break-up time, McMonnies survey, goblet cell density, and meibomian gland assessment. Conventional bacterial culture and broad-range 16S rDNA PCR, cloning, and DNA sequencing were used for bacterial identification. Repeated sampling was performed in a subset of subjects over a 3-month period. The association between goblet cell loss and bacterial counts in a subgroup of subjects was assessed. RESULTS. Most of the bacteria identified by culture were coagulase negative staphylococci, whereas molecular methods demonstrated a considerable number of additional bacteria. Atypical ocular surface bacteria including Rhodococcus erythropolis, Klebsiella oxytoca, and Erwinia sp., were identified in cases of overt inflammation and, surprisingly, on the normal ocular surface. The same bacteria remained on the ocular surface after repeated sampling. Increased bacterial flora was associated with reduced goblet cell density. CONCLUSIONS. Molecular analysis revealed a diverse ocular surface bacterial population. In addition to the normal flora, various potentially pathogenic bacteria were identified. The detection of known pathogens in both normal and dry eyes, with minimal signs of infection, presents a diagnostic dilemma. It remains unknown whether their presence is associated with inflammation and reduced goblet cell density or whether they adversely affect the ocular surface predisposing it to abnormal microbial colonization. In the absence of overt clinical infection, it is unknown whether such results should prompt intervention with therapy.",
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Ocular pathogen or commensal: A PCR-based study of surface bacterial flora in normal and dry eyes. / Graham, Joanna E.; Moore, Johnny; Jiru, Xu; Moore, John E.; Goodall, Edward A.; Dooley, James S. G.; Hayes, Velma; Dartt, Darlene A.; Downes, Stephen; Moore, Tara.

In: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, Vol. 48, No. 12, 01.12.2007, p. 5616-5623.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Ocular pathogen or commensal: A PCR-based study of surface bacterial flora in normal and dry eyes

AU - Graham, Joanna E.

AU - Moore, Johnny

AU - Jiru, Xu

AU - Moore, John E.

AU - Goodall, Edward A.

AU - Dooley, James S. G.

AU - Hayes, Velma

AU - Dartt, Darlene A.

AU - Downes, Stephen

AU - Moore, Tara

PY - 2007/12/1

Y1 - 2007/12/1

N2 - PURPOSE. To compare the bacterial population of the ocular surface of normal and dry eye subjects using conventional culture and 16S rDNA PCR. METHODS. Ninety-one subjects were classified as normal (n = 57) or dry eye (n = 34) by using tear break-up time, McMonnies survey, goblet cell density, and meibomian gland assessment. Conventional bacterial culture and broad-range 16S rDNA PCR, cloning, and DNA sequencing were used for bacterial identification. Repeated sampling was performed in a subset of subjects over a 3-month period. The association between goblet cell loss and bacterial counts in a subgroup of subjects was assessed. RESULTS. Most of the bacteria identified by culture were coagulase negative staphylococci, whereas molecular methods demonstrated a considerable number of additional bacteria. Atypical ocular surface bacteria including Rhodococcus erythropolis, Klebsiella oxytoca, and Erwinia sp., were identified in cases of overt inflammation and, surprisingly, on the normal ocular surface. The same bacteria remained on the ocular surface after repeated sampling. Increased bacterial flora was associated with reduced goblet cell density. CONCLUSIONS. Molecular analysis revealed a diverse ocular surface bacterial population. In addition to the normal flora, various potentially pathogenic bacteria were identified. The detection of known pathogens in both normal and dry eyes, with minimal signs of infection, presents a diagnostic dilemma. It remains unknown whether their presence is associated with inflammation and reduced goblet cell density or whether they adversely affect the ocular surface predisposing it to abnormal microbial colonization. In the absence of overt clinical infection, it is unknown whether such results should prompt intervention with therapy.

AB - PURPOSE. To compare the bacterial population of the ocular surface of normal and dry eye subjects using conventional culture and 16S rDNA PCR. METHODS. Ninety-one subjects were classified as normal (n = 57) or dry eye (n = 34) by using tear break-up time, McMonnies survey, goblet cell density, and meibomian gland assessment. Conventional bacterial culture and broad-range 16S rDNA PCR, cloning, and DNA sequencing were used for bacterial identification. Repeated sampling was performed in a subset of subjects over a 3-month period. The association between goblet cell loss and bacterial counts in a subgroup of subjects was assessed. RESULTS. Most of the bacteria identified by culture were coagulase negative staphylococci, whereas molecular methods demonstrated a considerable number of additional bacteria. Atypical ocular surface bacteria including Rhodococcus erythropolis, Klebsiella oxytoca, and Erwinia sp., were identified in cases of overt inflammation and, surprisingly, on the normal ocular surface. The same bacteria remained on the ocular surface after repeated sampling. Increased bacterial flora was associated with reduced goblet cell density. CONCLUSIONS. Molecular analysis revealed a diverse ocular surface bacterial population. In addition to the normal flora, various potentially pathogenic bacteria were identified. The detection of known pathogens in both normal and dry eyes, with minimal signs of infection, presents a diagnostic dilemma. It remains unknown whether their presence is associated with inflammation and reduced goblet cell density or whether they adversely affect the ocular surface predisposing it to abnormal microbial colonization. In the absence of overt clinical infection, it is unknown whether such results should prompt intervention with therapy.

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DO - 10.1167/iovs.07-0588

M3 - Article

VL - 48

SP - 5616

EP - 5623

JO - Investigative Ophthalmology and Visual Science

T2 - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 12

ER -