9 results on '"Tseng, Shin-Ling"'
Search Results
2. A dot hybridization assay for the diagnosis of bacterial keratitis
- Author
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Fang, Po-Chiung, Chien, Chun-Chih, Yu, Hun-Ju, Ho, Ren-Wen, Tseng, Shin-Ling, Lai, Yu-Hsuan, and Kuo, Ming-Tse
- Subjects
Keratitis ,Male ,Bacteria ,Humans ,Nucleic Acid Hybridization ,Female ,Middle Aged ,Corneal Ulcer ,Bacterial Load ,Eye Infections, Bacterial ,Research Article - Abstract
Purpose To evaluate a bacterial dot hybridization (BDH) assay for the diagnosis of bacterial keratitis (BK). Methods Sixty-one qualified corneal scrapings from 61 patients with suspected microbial keratitis were collected consecutively and prospectively. Among the 61 patients, 16 cases were BK and 45 cases were non-BK, including fungal keratitis, viral keratitis, parasitic keratitis, and non-microbial keratitis. Molecular diagnosis of BK in these corneal scrapes was performed using the BDH assay with three universal bacterial probes (PB1, PB2, and PB3) and three genus-specific probes (Aci, Klb, and Psu) to detect Acinetobacter, Klebsiella, and Pseudomonas, respectively. Signals were standardized after grayscale image transformation for objective validation using receiver operating characteristic (ROC) curves. Results The standardized intensities for the three universal probes differed statistically significantly between the BK group and the non-BK group. Based on the ROC curves, the sensitivities of PB1, PB2, and PB3 were 81.3%, 81.3%, and 93.8%, and the specificities were 71.1%, 88.9%, and 91.1%, respectively. The sensitivity and specificity of the Psu probe were 92% and 100%, respectively, while those of the Aci and Klb probes could not be estimated because there were no BK cases caused by Acinetobacter spp. or Klebsiella spp. Conclusions The BDH assay is an effective molecular approach to improve the diagnosis of BK. Because the bias from bacterial contamination on the ocular surface can be minimized with signal standardization, the assay has the potential to be adopted for routine clinical practice.
- Published
- 2017
3. PCR analysis for assessment of bacterial bioburden in orthokeratology lens cases
- Author
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Lo, Jung, Fang, Po-Chiung, Chien, Chun-Chih, Hsiao, Chang-Chun, Tseng, Shin-Ling, Lai, Yu-Hsuan, and Kuo, Ming-Tse
- Subjects
DNA, Bacterial ,Male ,Adolescent ,Bacteria ,Contact Lenses ,Bacterial Physiological Phenomena ,Polymerase Chain Reaction ,Biofilms ,Product Packaging ,Equipment Contamination ,Humans ,Female ,Prospective Studies ,Child ,Orthokeratologic Procedures ,Research Article - Abstract
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
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- 2016
4. Bacterial Bioburden Decrease in Orthokeratology Lens Storage Cases After Forewarning: Assessment by the DNA Dot Hybridization Assay
- Author
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Fang, Po-Chiung, primary, Lo, Jung, additional, Chang, Tsung C., additional, Chien, Chun-Chih, additional, Hsiao, Chang-Chun, additional, Tseng, Shin-Ling, additional, Lai, Yu-Hsuan, additional, and Kuo, Ming-Tse, additional
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- 2017
- Full Text
- View/download PDF
5. A Multiplex Dot Hybridization Assay For Detection and Differentiation ofAcanthamoebaand Herpes Keratitis
- Author
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Kuo, Ming-Tse, primary, Fang, Po-Chiung, additional, Yu, Hun-Ju, additional, Chao, Tsae-Ling, additional, Chien, Chun-Chih, additional, Chen, Shun-Hua, additional, Wang, Jen-Ren, additional, Tseng, Shin-Ling, additional, Lai, Yu-Hsuan, additional, Hsiao, Chang-Chun, additional, and Chang, Tsung C., additional
- Published
- 2016
- Full Text
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6. Microbial Bioburden of Orthokeratology Contact Lens Care System
- Author
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Lo, Jung, primary, Kuo, Ming-Tse, additional, Chien, Chun-Chih, additional, Tseng, Shin-Ling, additional, Lai, Yu-Hsuan, additional, and Fang, Po-Chiung, additional
- Published
- 2016
- Full Text
- View/download PDF
7. A DNA dot hybridization model for molecular diagnosis of parasitic keratitis.
- Author
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Huang FC, Hsieh HY, Chang TC, Su SL, Tseng SL, Lai YH, and Kuo MT
- Subjects
- Acanthamoeba genetics, Acanthamoeba Keratitis parasitology, Corneal Ulcer diagnosis, Corneal Ulcer microbiology, DNA, Fungal genetics, Eye Infections, Fungal diagnosis, Eye Infections, Fungal microbiology, Eye Infections, Parasitic parasitology, False Negative Reactions, Humans, Microsporidia genetics, Microsporidiosis diagnosis, Microsporidiosis microbiology, Molecular Diagnostic Techniques, Oligonucleotide Probes chemistry, Polymerase Chain Reaction, Predictive Value of Tests, Prospective Studies, RNA, Ribosomal genetics, RNA, Ribosomal, 18S genetics, Ribosome Subunits, Small genetics, Sensitivity and Specificity, Acanthamoeba Keratitis diagnosis, DNA, Protozoan genetics, Eye Infections, Parasitic diagnosis, Nucleic Acid Hybridization methods
- Abstract
Purpose: Developing a DNA dot hybridization model for diagnosing parasitic keratitis., Methods: Newly designed oligonucleotide probes for detecting Acanthamoeba and microsporidia were tested with target reference strains of Acanthamoeba (n = 20) and microsporidia (n = 3), and non-target microorganisms, including bacteria (n = 20) and fungi (n = 20). These probes, which had passed the preliminary tests, were then assembled as a parasite dot hybridization (PDH) model for assessing 33 clinical samples from patients with clinically suspected Acanthamoeba and microsporidia keratitis, including eight positives for Acanthamoeba , 13 positives for microsporidia, and 12 negatives for both pathogens., Results: Two probes for detecting Acanthamoeba and two for detecting microsporidia passed the tests using target and non-target strains and then were assembled in the PDH model. For clinical samples, one Acanthamoeba -positive sample (proved with pathology) was falsely negative according to the PDH assay. The sensitivity and specificity of the PDH assay for diagnosing Acanthamoeba keratitis were 87.5% and 100%, respectively, while the sensitivity and specificity for diagnosing microsporidia keratitis were 100%. The infectious agent of all clinical samples of microsporidia keratitis was identified as Vittaforma corneae with DNA sequencing, while those of Acanthamoeba keratitis were caused by four species of Acanthamoeba , with Acanthamoeba castellanii found in four samples (50%, 4/8)., Conclusions: The PDH model has the potential to be a molecular assay for diagnosing Acanthamoeba and microsporidia keratitis. However, a prospective clinical study might be needed before the model is adopted in routine clinical practice.
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- 2017
8. A Multiplex Dot Hybridization Assay For Detection and Differentiation of Acanthamoeba and Herpes Keratitis.
- Author
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Kuo MT, Fang PC, Yu HJ, Chao TL, Chien CC, Chen SH, Wang JR, Tseng SL, Lai YH, Hsiao CC, and Chang TC
- Subjects
- Cornea parasitology, Cornea virology, Diagnosis, Differential, Humans, Limit of Detection, Multiplex Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Acanthamoeba, Acanthamoeba Keratitis diagnosis, Keratitis, Herpetic diagnosis, Nucleic Acid Hybridization methods, Simplexvirus
- Abstract
Purpose: We verified a multiplex dot hybridization (MDH) assay for the rapid detection and differentiation of Acanthamoeba keratitis (AK) and herpes simplex keratitis (HSK)., Methods: Molecular detection of Acanthamoeba and herpes simplex virus in corneal scrapes was performed with the MDH assay and standard diagnostic methods. The experimental group included corneal scrapes (n = 33) from patients with culture- or pathology-confirmed AK (n = 15) and real-time PCR-confirmed HSK (n = 16). The control group included 50 samples from cases of bacterial keratitis (n = 15), fungal keratitis (n = 15), and initially presumed AK (n = 5) or HSK (n = 17) which finally were excluded by culture for Acanthamoeba or by real-time PCR for herpes simplex virus, respectively. Discrepant results between methods were resolved by DNA sequencing of the PCR amplicons., Results: After discrepant analysis, the sensitivity for the diagnosis of AK and HSK was both 93.3% by the MDH assay, while the specificity was 100% for the two types of keratitis. The turnaround time of MDH assay was within a working day using an already prepared array. Two false-negatives (one AK case and one HSK case) were obtained by the MDH assay., Conclusions: The MDH assay could effectively prevent missed or delayed diagnosis of AK and HSK and has a potential to be adopted in routine clinical practice if the test is commercialized.
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- 2016
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9. A DNA dot hybridization model for assessment of bacterial bioburden in orthokeratology lens storage cases.
- Author
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Kuo MT, Chien CC, Lo J, Hsiao CC, Tseng SL, Lai YH, Fang PC, and Chang TC
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- Adolescent, Bacteria genetics, Bacterial Physiological Phenomena, Biofilms growth & development, Child, Colony Count, Microbial, DNA Probes chemistry, Female, Humans, Male, Polymerase Chain Reaction, Prospective Studies, Bacteria isolation & purification, Contact Lenses microbiology, DNA, Bacterial analysis, Equipment Contamination, Nucleic Acid Hybridization methods, Orthokeratologic Procedures instrumentation, Product Packaging
- Abstract
Purpose: The aim of this study was to evaluate a DNA dot hybridization assay (DHA) for assessing bacterial bioburden in orthokeratology lens (OK) storage cases., Methods: Forty-one OK wearers participated in this study. The dot hybridization assay was used to assess the bacterial bioburden of OK after removal and 6-hour soaking in a storage case. Signals of the DHA were standardized after gray image transformation. The correlations between the hybridization intensities of three universal bacteria probes (BP1, BP2, and BP3) and bacterial bioburden determined by culture (colony forming units per milliliter) was analyzed by Pearson's correlation coefficient and receiver operating characteristic plots. In addition, three genus-specific probes for Pseudomonas, Acinetobacter, and Klebsiella were used to detect potentially hazardous bacterial contamination regardless of bacterial viability status., Results: Among the three universal probes, there were good correlations between probe BP2 (r2 = 0.31, P = 9.5 × 10(-5)) and probe BP3 (r2 = 0.35, P = 3.1 × 10(-5)) with bacterial bioburden, but no correlation was found between probe BP1 and bacterial bioburden (r2 = 0.04, P = 0.11). In 41 samples, one was Pseudomonas-positive by both DHA and culture, while 10 were Pseudomonas-positive by DHA but negative by culture. Furthermore, nine samples tested positive for Acinetobacter (n = 7) and Klebsiella (n = 2) by DHA only., Conclusions: The dot hybridization assay provides a novel way to assess the bacterial bioburden of OK storage cases. Lens care quality can be assessed with universal bacteria probes, while potentially hazardous bacterial contamination can be traced with genus-specific probes., (Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.)
- Published
- 2014
- Full Text
- View/download PDF
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