Evaluation of real-time PCR method for rapid diagnosis of brucellosis with different clinical manifestations.

In this study, we tested the advantages of TaqMan real time PCR technique and compare it to conventional methods using serum samples from patients with different clinical forms of brucellosis. A total of 50 patients were included in the study. Blood culture using BACTEC 9240 system, Standard Wright's tube agglutination, and real time PCR methods were used. Control blood samples from 30 people with no history of brucellosis or exposure to Brucella spp. were examined too. Serological assay was positive for 49 patients (98%). Forty-four (88%) of the 50 patients had a positive PCR result, whereas Brucella spp were isolated from blood cultures of 18 patients (36%). STA test was all positive for focal brucellosis. Real time PCR test was positive in 9 patients with focal disease (90%), whereas blood culture was positive only in 4 patients (40%). The sensitivity, specificity, positive and negative predictive values of the real time PCR method were calculated as 88%, 100%, 100%, and 83%, respectively. Our results suggest that the high sensitivity and specificity of real time PCR method make it a useful tool for diagnosis of brucellosis with different clinical manifestations.

[The relationship between paraoxanase gene Leu-Met (55) and Gln-Arg (192) polymorphisms and coronary artery disease]. / Paraoksonaz geninde Leu-Met (55) ve Gln-Arg (192) polimorfizmleri ile koroner arter hastaligi arasindaki iliski.

OBJECTIVES: Paraoxonase (PON1) is a high-density lipoprotein (HDL)-associated esterase that hydrolyses lipoperoxides. PON1 serves as a protective factor against oxidative modification of LDL, suggesting that it may play an important role in the prevention of atherosclerotic process. Research has focused on two polymorphisms: leucine (L allele) to methionine (M allele) substitution at codon 55, and glutamine (A allele) to arginine (B allele) substitution at codon 192. STUDY DESIGN: We examined amino acid changes at codon 55 and 192 in the PON1 gene by polymerase chain reaction and using restriction enzymes in 120 patients (92 men, 28 women; mean age 48.2+/-4.3 years) with premature coronary artery disease (CAD) and in 102 healthy subjects (80 men, 22 women; mean age 46.8+/-5.2 years) with no history of CAD and a normal electrocardiogram. RESULTS: Distribution of genotypes in the patient and control groups at codon 55 were 6.7% and 4.9% for MM, 46.7% and 29.4% for LM, 46.7% and 65.7% for LL, respectively. The frequency of genotypes at codon 192 were as follows: 4.2% and 2% for RR, 40% and 35.3% for QR, and 55.8% and 62.8% for QQ, respectively. While the frequency of PON1 55M allele was higher in the CAD group (0.3 vs. 0.2), PON1 192R allele frequency did not differ (p>0.05). There was a significant relationship between the PON1 M/L55 polymorphism and CAD (p=0.017), whereas the R/Q192 polymorphism was not associated with CAD (p=0.445). CONCLUSION: These data suggest that the PON1 M/L55 polymorphism shows a significant relationship with CAD and the Q/R192 polymorphism is not a major risk factor causing susceptibility to CAD in our population.