DETEKSI DAN VARIASI GENETIK LEPTOSPIRA PADA TIKUS, AIR DAN TANAH DI KABUPATEN BANTUL DIY

HADI SUMANTA, Prof. dr. Hari Kusnanto, SU, Dr.PH; dr. Tri Wibawa, Sp.MK, Ph.D.; Prof. Dr. Drs. Suwarno Hadisusanto, S.U

2015 | Disertasi | S3 Kedokteran Umum

Latar belakang: Upaya pencegahan leptospirosis di Kabupaten Bantul sudah dilakukan. Namun, kejadian leptospirosis selalu ditemukan sepanjang tahun. Itu terjadi karena pencegahan dan pengendalian belum berorientasi pada Leptospira sebagai sumber infeksi. Tikus, air dan tanah merupakan sumber Leptospira yang menyebabkan leptospirosis. Keberadaan Leptospira di lingkungan dapat dideteksi dengan RT-PCR, dan sebaran Leptospira dianalisis dengan spasial. Tujuan: Penelitian ini bertujuan untuk menganalisis Leptospira di tikus, air dan tanah, mendeteksi variasi gen 16S rRNA Leptospira dengan PCR, menganalisis hubungan keberadaan Leptospira di tikus, air dan tanah dengan faktor lingkungan, menganalisis sebaran clustering, dan prediksi sebaran tikus, air dan tanah sebagai sumber Leptospira. Metode:Penelitian ini bersifat observasional menggunakan rancangan cross sectional. Sampel tikus sebanyak 99 ekor, air 119 sampel dan tanah 150 sampel diambil di sekitar tempat tinggal penderita leptospirosis. Leptospira dideteksi dengan teknik RT-PCR, menggunakan primer yang dapat mendeteksi gen 16S rRNA Leptospira. Amplifikasi gen 16S rRNA menggunakan PCR, kemudian amplikon di sequencing. Analisis variasi gen menggunakan MEGA 6.2. Analisis univariat dan bivariat menggunakan STATA. Analisis sebaran dan clutering menggunakan SatScan, sedangkan prediksi sebaran Leptospira menggunakan CrimeStat. Hasil dan Simpulan: Ditemukan Leptospira patogen pada tikus dengan proporsi 25.25%, tanah 42.67% dan air 42.68%. Gen 16S rRNA Leptospira patogenik dari sampel tikus memiliki kemiripan dengan sequence gene bank sampel dari manusia, tikus, ternak dan lingkungan. Keberadaan Leptospiradi tikus tidak berhubungan dengan faktor vegetasi, sampah, kandang ternak, genangan air, suhu, kelembaban dan pH. Sebaran sumber Leptospira membentuk clustering dan diprediksi menyebar keluar wilayah Kabupaten Bantul.

Background: Incidence of leptospirosis remain high throughout the year. There were several programs have been to prevent distribution. Unfortunately, the programs was not successful, due to unfocused orientation of sources of infection, mainly rats, water and soil. RT-PCR has been used widely to detect Leprospira existence, whereas spatial analysis used for Sprira distribution. This study aims were to identify and analyze distribution of Leptospira in rat, water and soil. Objectives: The objectives of this research were identifying Leptospira in rats, water and soil, and determined variation in 16S rRNA gene by RT-PCR Leptospira; Analyzing the relationship existence Leptospira in rats, water and land with environmental factors; Analyzing the distribution of clustering, and prediction of the distribution of rat, water and soil as a source of Leptospira. Method: the study was a cross sectional design. Rats, water and soil as sample materials were taken from the surrounding leptospirosis patients. Leptospira was determined using RT-PCR technic, mainly using the primer that can detect 16S rRNA Leptospira gen. 16S rRNA gene amplification using PCR, and then amplicons was sequenced. MEGA 6.2 was used to Analysis of gene variations, STATA to analyze. Univariate analyzes and bivariate, whereas distribution and clustering were analyzed using SatScan, and prediction distribution of Leptospira using CrimStat. Resultsand Conclusion: the proportion of Leptospira pathogens in rats were 25.25%, soil 42.67%, and water 42.68% respectively. The 16S rRNA genes of pathogenic Spira in rat were similar with gene bank sequences from sample of human, mice, animals and environment. The existence of Spira are not associated with vegetation, litter, cattle sheds, poolsof water, temperature, humidity and pH. Distribution of rats, water and soil as a source of Leptospira is forming a cluster, and we predict that it has been spread out of Bantul Regency.

Kata Kunci : Leptospira, deteksi,variasi genetik, clustering, prediksi.