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1.
Clin Ophthalmol ; 6: 1977-80, 2012.
Article in English | MEDLINE | ID: mdl-23226003

ABSTRACT

We report an unusual case of iris rubeosis and hyphema caused by chemical injury due to household detergent. A 74-year-old man with a 15-year history of diabetes mellitus was refilling a container with household detergent at home. He splashed the detergent in his eyes. Slit-lamp examination revealed extensive epithelial damage to the left eye, leading to a persistent corneal epithelial defect. We used a bandage soft contact lens with levofloxacin eye drops as concomitant therapy in order to promote healing. However, a strain of fluoroquinolone-resistant Corynebacterium colonized the eye, so that the corneal ulcer eventually became severe. Use of the bandage soft contact lens was discontinued. His antimicrobial agent was changed to cefmenoxime, a drug to which fluoroquinolone-resistant Corynebacterium is sensitive, and topical instillation of autologous serum subsequently promoted improvement of the ulcer. On day 38 after injury, iris rubeosis led to hyphema and ghost cell glaucoma. With improvement of his corneal epithelial defect, the iris rubeosis and hyphema regressed and his visual acuity improved to 20/25 on the left eye. To the best of our knowledge, this is the first report of a case resulting in severe complications due to chemical injury by a neutral detergent. Ophthalmologists should be aware that corneal epithelial damage may become prolonged in elderly patients with diabetes, and unexpectedly severe when wearing bandage soft contact lens, with infection of Corynebacterium resistant to fluoroquinolones, even if the chemical agent is a neutral detergent.

2.
J Clin Microbiol ; 42(9): 4284-92, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15365024

ABSTRACT

Vibrio parahaemolyticus strains carrying the thermostable direct hemolysin (TDH) tdh gene, the TDH-related hemolysin (trh) gene, or both genes are considered virulent strains. We previously demonstrated that the transcription-reverse transcription concerted (TRC) method could be used to quantify the amount of mRNA transcribed from the tdh gene by using an automated detection system. In this study, we devised two TRC-based assays to quantify the mRNAs transcribed from the trh1 and trh2 genes, the two representative trh genes. The TRC-based detection assays for the tdh, trh1, and trh2 transcripts could specifically and quantitatively detect 10(3) to 10(7) copies of the corresponding calibrator RNAs. We examined by the three TRC assays the total RNA preparations extracted from 103 strains of Vibrio parahaemolyticus carrying the tdh, trh1, or trh2 gene in various combinations. The tdh, trh1, and trh2 mRNAs in the total RNA preparations were specifically quantified, and the time needed for detection ranged from 9 to 19 min, from 14 to 18 min, and from 9 to 12 min, respectively. The results showed that this automated TRC assays could detect the tdh, trh1, and trh2 mRNAs specifically, quantitatively, and rapidly. The relative levels of TDH determined by the immunological method and that of tdh mRNA determined by the TRC assays for most tdh-positive strains correlated. Interestingly, the levels of TDH produced from the strains carrying both tdh and trh genes were lower than those carrying only the tdh gene, whereas the levels of mRNA did not significantly differ between the two groups.


Subject(s)
Hemolysin Proteins/genetics , RNA, Bacterial/genetics , RNA, Messenger/genetics , Vibrio parahaemolyticus/genetics , Automation/methods , Base Sequence , DNA Primers , Geography , Humans , Models, Molecular , Molecular Sequence Data , Nucleic Acid Conformation , Oligonucleotide Probes , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction/methods , Transcription, Genetic/genetics , Vibrio parahaemolyticus/classification , Vibrio parahaemolyticus/isolation & purification
3.
Anal Biochem ; 314(1): 77-86, 2003 Mar 01.
Article in English | MEDLINE | ID: mdl-12633605

ABSTRACT

We developed a completely homogeneous and isothermal method of detecting RNA sequences and demonstrated ultrarapid and direct quantification of pathogenic gene expression with high sensitivity. The assay is based on performing isothermal RNA sequence amplification in the presence of our novel DNA probe, an intercalation activating fluorescence DNA probe, and measuring the fluorescence intensity of the reaction mixture. When detecting mecA gene expression of methicillin-resistant Staphylococcus aureus, we quantified starting copies ranging from 10 to 10(7) copies within 10min. The primer sequences were designed to bind to secondary structure-free sites of the target RNA, which enabled a totally isothermal protocol to quantify mRNA specifically in a sample of existing genomic DNA. When we applied this to quantifying the expression of marker genes of Vibrio parahaemolyticus and Mycobacterium bovis BCG strain, the results correlated well with the viability of each bacterium. We also demonstrated monitoring Pab gene expression of M. bovis BCG during cultivation with antibiotics. The present method can potentially realize rapid antimicrobial susceptibility testing of slowly growing organisms, such as tuberculosis.


Subject(s)
DNA Probes/metabolism , Intercalating Agents/metabolism , Polymerase Chain Reaction/methods , Base Sequence , DNA Primers/metabolism , DNA, Bacterial/genetics , Fluorescence , Fluorescent Dyes/metabolism , Gene Expression Regulation, Bacterial , Humans , Molecular Sequence Data , Molecular Structure , Mycobacterium bovis/genetics , RNA, Bacterial/analysis , RNA, Bacterial/genetics , Sensitivity and Specificity , Staphylococcus aureus/genetics , Temperature , Vibrio parahaemolyticus/genetics
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