Background: It is important to diagnosis and properly treat patients with diarrhea is having a highly sensitive and specific technique to rapidly identify the caused bacteria, especially Diarrheagenic Escherichia Coli (DEC) isolates. Objectives: To develop and complete the bacterial DNA extraction procedure and optimize the DNA concentration for multiplex PCR for DEC. Materials and method: 7 reference strains of DEC and 10 fecal samples taken randomly were tested using DNA extraction and PCR techniques. Results. A bacterial DNA extraction procedure has been developed and optimized. This is a simple process and does not require expensive equipment. The test result is available after 90 \u2013 100 minutes. The minimum DNA content required for PCR to give positive results is 100ng per reaction. Conclusion: The development and completion of DNA extraction procedure plays an important role in early detection of DEC in fecal samples and serves as a base for further research on diarrheagenic bacteria.
DNA extraction ; PCR ; Escherichia coli

OBJECTIVETo investigate the association of polymorphisms of nucleotide excision repair genes and chromosomal damage in peripheral blood lymphocytes among coke-oven workers.

METHODSThe genotypes of ERCC1 C19007T, ERCC2 C22541A, ERCC2 G23591A, ERCC2 A35931C, ERCC4 T30028C, ERCC5 G3507C and ERCC6 A3368G among 140 coke-oven workers and 66 non-coke-oven controls were determined by PCR-PFLP methods. Chromosomal damage was detected by cytokinesis-block micronucleus (CBMN) assay.

RESULTSMultivariate analysis of covariance revealed that in coke-oven workers, the ERCC1 19007 CC genotype exhibited significantly higher CBMN frequency [(1.05 +/- 0.68)%] than did the CT [(0.81 +/- 0.66)%] (P = 0.01) or TT [(0.66 +/- 0.37)%] (P = 0.05) or CT + TT genotypes [(0.75 +/- 0.63)%] (P = 0.004). For the ERCC6 A3368G polymorphism, AA genotype exhibited significantly higher CBMN frequency [(1.00 +/- 0.69)%] than did the AG [(0.67 +/- 0.42)%] (P = 0.05) or AG + GG genotypes [(0.66 +/- 0.41)%] (P = 0.02). Stratification analysis found the significant association between the two polymorphisms, ERCC1 C19007T and ERCC6 A3368G, and the CBMN frequencies were most pronounced in older workers. In addition, for the polymorphism of ERCC2 G23591A, GA carriers had significantly higher CBMN frequencies [(1.40 +/- 0.63)%] than those GG carriers [(0.98 +/- 0.59)%] (P = 0.01) in older workers.

CONCLUSIONSOur results suggested that polymorphisms of ERCC1 C19007T, ERCC6 A3368G and ERCC2 G23591A were associated with the CBMN frequencies in coke-oven workers.

Adult ; Alcohol Drinking ; Coke ; Comet Assay ; DNA Damage ; DNA Repair ; genetics ; DNA Repair Enzymes ; genetics ; Dihydroxydihydrobenzopyrenes ; urine ; Extraction and Processing Industry ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lymphocytes ; Male ; Micronucleus Tests ; Middle Aged ; Occupational Exposure ; analysis ; Polymorphism, Genetic ; Smoking

A 37-year-old female presented to our hospital with a history of bleeding episodes (excessive bleeding after tooth extraction, gum bleeding, easy bruising, and excessive menstruation) and severe thrombocytopenia (2,000/µL). She had no family history of bleeding tendency or thrombocytopenia. No peripheral lymphadenopathy or splenomegaly was noted. The patient's white blood cell count was normal; hemoglobin was 9.7 g/dL. A peripheral blood smear showed markedly decreased platelets, with occasional giant or large platelets. Bone marrow examination found increased megakaryocytes. The patient also complained of hearing difficulty; a hearing test indicated sensory-neural hearing impairment. Her thrombocytopenia was refractory to treatment with glucocorticosteroids, intravenous gamma-globulin, and danazol. In the 13 years following her initial presentation, the patient required anti-hypertensive treatment, a hearing-aid for progressive hearing loss, and started maintenance kidney dialysis. Her clinical history of refractory thrombocytopenia, progressive hearing impairment, and renal failure suggested myosin heavy chain 9 gene-related congenital syndrome (Epstein syndrome), which was confirmed by the presence of a heterozygous deletion mutation, c.221_223del, (p.Lys74del) in peripheral leukocyte deoxyribonucleic acid.
Adult ; Bone Marrow Examination ; Danazol ; Dialysis ; DNA ; Female ; gamma-Globulins ; Gingiva ; Hearing ; Hearing Loss ; Hearing Loss, Sensorineural ; Hearing Tests ; Hemorrhage ; Humans ; Kidney ; Leukocyte Count ; Leukocytes ; Lymphatic Diseases ; Megakaryocytes ; Myosin Heavy Chains ; Renal Insufficiency ; Renal Insufficiency, Chronic ; Sequence Deletion ; Splenomegaly ; Thrombocytopenia* ; Tooth Extraction