An In-House Method for Molecular Monitoring of BCR-ABL.

OBJECTIVE: At present, there are a limited number of facilities in Turkey that can provide reliable real-time quantitative(RQ)-PCR BCR-ABL results. The present study aimed to test a cost-effective, in-house method of BCR-ABL quantification,including verification of the method by RQ-PCR validation tests. MATERIAL AND METHODS: BCR-ABL and ABL target sequences were cloned into pJET1.2 vectors, from whichcalibrators were prepared and used as templates in RQ-PCR reactions to generate standard curves. Dilutions of K562cells (representing an in vitro simulation of BCR-ABL transcript reduction) were analyzed. RESULTS: Standard curves were generated from calibrators. These curves were then used to calculate the BCR-ABL andABL copy numbers; in which linear BCR-ABL results were obtained. Repetitive experiments showed that our methodologywas able to detect 1 BCR-ABL positive cell from amnong 1x105 cells. CONCLUSION: The method described herein is suitable for implementation with any RQ-PCR instrument and/or kit forquantify BCR-ABL transcripts. CONFLICT OF INTEREST: None declared.

Microbial keratitis in West Anatolia, Turkey: a retrospective review.

PURPOSE: To analyze the clinical and microbiological characteristics of microbial keratitis in West Anatolia, Turkey during a 16-year period. METHODS: This is a retrospective study in which medical records were reviewed for all eyes with clinically diagnosed microbial keratitis presenting at the Izmir Ataturk Training and Research Hospital from January 1990 to December 2005. Risk factors, microbial isolations and clinical features were analyzed. RESULTS: Microbial keratitis was diagnosed for 620 eyes in 620 patients (250 women and 370 men; mean age: 54.13 years +/- 20.06). Pathogens from cultures were identified for 225 eyes (36.2%) and included Gram-negative (8.9%) and Gram-positive micro-organisms (68.8%) and fungi (22.3%). Staphylococcus epidermidis was the most commonly isolated micro-organism (26.6%), followed by S. aureus (24.4%), Streptococci pneumoniae (15.5%), fungi (22.3%), Pseudomonas species (6.6%), Enterobacter (2.2%) and Corynebacterium (2.2%). Ocular trauma was the most common predisposing factor (26.6%). Mean duration from onset of symptoms to diagnosis was 11.68 +/- 11.29 days. CONCLUSION: Staphylococci are the most commonly isolated microbial agents in cases of infectious keratitis in West Anatolia, Turkey. These findings will provide valuable background information and facilitate successful treatment in this region, and they will be useful as reference data for the particular population in this region as a basis for empirical antibiotic therapy.

Comparison of incidence of Demodex folliculorum on the eyelash follicule in normal people and blepharitis patients.

Demodex folliculorum has been incriminated in the development of blepharitis although much controversy persist. Some authors suggest that Demodex is a direct pathogen in chronic palpebral conditions while others consider the saprophyte to be innocuous to skin. We conducted a prospective study of eyelashes in 48 patients with blepharitis (totally 96 eyes) and 48 healthy persons (totally 96 eyes), searching for Demodex folliculorum and chronic blepharitis. Microscopy in immersion oil after storage in a moist chamber was performed. It is determined Demodex follicullorum in 11 of 37 (29.72%) patients with blepharitis, in one of 11 (9.09%) patient with blepharoconjunctivitis and in 2 of 48 (4.16%) persons in healthy control group. Staphylococcus aureus was cultivated in five patients and D. folliculorum was found two of them. The incidence of Demodex folliculorum was high in patients with blepharitis compared with normal controls. Incidence was high in males. Anti-Demodex treatment is indicated when the parasite is found.