Detection of sensitivity, PCR-RFLP metods, targeted spesific sequence in LipL32, OmpL1 and 16S rRNA gene regions for diagnosis leptospirosis

Leptospirosis is one of the most important zoonozis, seen in regions with temperate and rainy climate, including our region. Causing a wide clinical spectrum, clinical diagnosis of Leptospirosis is extremely difficult because only 10 % of cases with nonspecific symptoms can be seen. Most of the cases of Leptospirosis are either non-specific empirical treatment without diagnosis, or disease terminated with spontaneous recovery. However, with patients in the last icteric period, it is necessary to admit them to research hospitals, and evaluate them via non-specific and low sensitivity methods with blood and urine samples through dark field microscopy.This study is aimed at developing alternative methods for identifying deficiencies in conventional diagnosis methods, diagnostic kit based PCR-RFLP, which belong to the suspect patient clinical materials; a- to show existence leptosipira, b- identification of L.interrogans and L.biflexa types, with high sensitivity and specificity and can produce rapid results.A total of 47 human patients, 38 adults and 9 under of 18 age, were admitted to Ç.U. Faculty of Medicine and The Depertment of Infectious Disaease of Adana State Hospital of Health Ministery, and blood samples of 86 cattle belongs to Çukurova region which showed hematuria and icter symptomps, were investigated with 16S rRNA Nested-PCR RFLP, OmpL1 PCR and LipL32 nested-PCR tests, to create more sensitive, specific and rapid alternative diagnostic tests to determine the usability of DFM and MAT.While positivity rates in all samples analyzed in DFM methods, 22 human and 34 animal, sera appearance were obtained by Leptospira compatible images, the result of the MAT samples from 7 different L.interrogans strains and L.patoc strain were used as an antigen to evaluate, 21 human and 69 animal serum samples seropositivity were detected in at least 1/50 via antigen titration. The target sequences with in 16SrRNA gene were found to be positive in 92 patients, of which 77 belonged to the L.interrogans and 15 L.biflexa by using 16S rRNA-specific nested-PCR-RFLP which allows the separation of pathogenic and non-pathogenic strains. And the result of LipL32 gene specific primers PCR in 77 samples of all samples L.interrogans was found, and by using OmpL1 PCR which allows separation of the level of serotype to L.interrogans, 67 samples were found to be positiveL.hardjo have been identified as the most common serotypes in the region with rate is 17% including 2,1% of people and 25,6% of animal samples and except for this serotype, L.grippotyphosa, L.bratislava and L.ichteriohaemmorhagia respectively have been seen as the serotypes identified.