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Gene Synthesis> | All extracted DNAs of rodent's tissues were selected for Crithidia detection, using minicircle kDNA sequences of internal transcribed spacer (ITS) gene. The specific primers were designed by GenScript online PCR primers designs tool by using gene bank information (Unpublished document). Each 25-μL reaction mixture (total volume) contained 12 μL Ampliqon master mix buffer, 1 μL of each primers of 5′-TCCATGTGCGAGGACAACGTGCT- 3′and 3′-CGCGTCGTTGATGAAGTCGCT-5′ (with 10 pico mol concentrations), 5 μL DNA sample and 6 μL double Distilled water. Eppendorf Master-thermocycler programmed for an initial temperature of 94 °C for 5 min (one cycle), followed by temperatures of 94 °C, 55 °C, 72 °C, for 30 s, 1 min, and 1.5 min, respectively (30 cycles), and continued by a final temperature of 72 °C for 5 min (one cycle). Afterward, 5 μL of PCR products were used for electrophoresis assay. Reference strain of Crithidia fasciculata was used and a band of 800 bp would have shown the existence of Crithidia in the tested smears. | Get A Quote |
Objective: To explore the co-detection of natural infection of Trypanosomatidae parasites such as Leishmania and Crithidia in reservoir hosts of leishmaniasis. Methods: Rodent populations were monitored in two endemic foci of cutaneous leishmaniasis of Fars province, southern Iran from March to October 2016. Rodents were trapped alive in several parts of Shiraz and Kharameh cities. Afterwards, their organs were prepared for detection of Leishmania and Crithidia species by molecular, microscopic, and culture methods. Results: Totally, 115 rodents of five species; Tatera indica (T. indica) (85), Rattus rattus (12), Meriones libycus (9), Mus musculus (7), and Rattus norvegicus(2), were trappe... More