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PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases

PURPOSE: To develop a PCR gel analysis method for assessing the bacterial bioburden in orthokeratology contact lens (OK) case fluid determined by culture. METHODS: A prospective study with the participation of 41 OK wearers (20 girls, 21 boys) was performed. The mean OK-wearing experience was 3.5±1....

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Autores principales: Lo, Jung, Fang, Po-Chiung, Chien, Chun-Chih, Hsiao, Chang-Chun, Tseng, Shin-Ling, Lai, Yu-Hsuan, Kuo, Ming-Tse
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Vision 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734148/
https://www.ncbi.nlm.nih.gov/pubmed/26900321
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author Lo, Jung
Fang, Po-Chiung
Chien, Chun-Chih
Hsiao, Chang-Chun
Tseng, Shin-Ling
Lai, Yu-Hsuan
Kuo, Ming-Tse
author_facet Lo, Jung
Fang, Po-Chiung
Chien, Chun-Chih
Hsiao, Chang-Chun
Tseng, Shin-Ling
Lai, Yu-Hsuan
Kuo, Ming-Tse
author_sort Lo, Jung
collection PubMed
description PURPOSE: To develop a PCR gel analysis method for assessing the bacterial bioburden in orthokeratology contact lens (OK) case fluid determined by culture. METHODS: A prospective study with the participation of 41 OK wearers (20 girls, 21 boys) was performed. The mean OK-wearing experience was 3.5±1.9 years. PCR was used to assess the bacterial bioburden (colony-forming units per milliliter) of OK after removal and soaking in the storage case for 6 h. The signal intensity of the PCR bands was analyzed after grayscale image transformation. The difference (cPCR-d) and ratio (cPCR-r) between a PCR signal and its background were used as two standardized indices of PCR signals. The association between the two indices of the PCR signals and the bacterial bioburden determined by culture were analyzed with Pearson’s correlation coefficient (r) and receiver operating characteristic (ROC) plots. RESULTS: At least one microbe was isolated from the OK lens case from 38 of the 41 subjects. Both cPCR-d and cPCR-r showed strong correlations with the bacterial bioburden (r>0.7, p<0.0001). ROC analysis enabled good determination of the cutoff values for the two PCR indices with acceptable sensitivity and specificity (78−89%) to assess the degree of bacterial contamination. CONCLUSIONS: The high microbial contamination rate of the OK lens cases revealed the general inappropriate lens care by OK wearers. PCR analysis provides an alternative and rapid method for assessing the bacterial bioburden of OK lens cases, and these results should serve as a warning to OK wearers to follow appropriate lens care procedures to prevent infection.
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spelling pubmed-47341482016-02-19 PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases Lo, Jung Fang, Po-Chiung Chien, Chun-Chih Hsiao, Chang-Chun Tseng, Shin-Ling Lai, Yu-Hsuan Kuo, Ming-Tse Mol Vis Research Article PURPOSE: To develop a PCR gel analysis method for assessing the bacterial bioburden in orthokeratology contact lens (OK) case fluid determined by culture. METHODS: A prospective study with the participation of 41 OK wearers (20 girls, 21 boys) was performed. The mean OK-wearing experience was 3.5±1.9 years. PCR was used to assess the bacterial bioburden (colony-forming units per milliliter) of OK after removal and soaking in the storage case for 6 h. The signal intensity of the PCR bands was analyzed after grayscale image transformation. The difference (cPCR-d) and ratio (cPCR-r) between a PCR signal and its background were used as two standardized indices of PCR signals. The association between the two indices of the PCR signals and the bacterial bioburden determined by culture were analyzed with Pearson’s correlation coefficient (r) and receiver operating characteristic (ROC) plots. RESULTS: At least one microbe was isolated from the OK lens case from 38 of the 41 subjects. Both cPCR-d and cPCR-r showed strong correlations with the bacterial bioburden (r>0.7, p<0.0001). ROC analysis enabled good determination of the cutoff values for the two PCR indices with acceptable sensitivity and specificity (78−89%) to assess the degree of bacterial contamination. CONCLUSIONS: The high microbial contamination rate of the OK lens cases revealed the general inappropriate lens care by OK wearers. PCR analysis provides an alternative and rapid method for assessing the bacterial bioburden of OK lens cases, and these results should serve as a warning to OK wearers to follow appropriate lens care procedures to prevent infection. Molecular Vision 2016-01-14 /pmc/articles/PMC4734148/ /pubmed/26900321 Text en Copyright © 2016 Molecular Vision. http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited, used for non-commercial purposes, and is not altered or transformed.
spellingShingle Research Article
Lo, Jung
Fang, Po-Chiung
Chien, Chun-Chih
Hsiao, Chang-Chun
Tseng, Shin-Ling
Lai, Yu-Hsuan
Kuo, Ming-Tse
PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title_full PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title_fullStr PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title_full_unstemmed PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title_short PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
title_sort pcr analysis for assessment of bacterial bioburden in orthokeratology lens cases
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734148/
https://www.ncbi.nlm.nih.gov/pubmed/26900321
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