en انگل شناسی Parasitology گزارش کوتاه Brief Report <span style="font-size:12px;"><span style="font-family:Times New Roman;"><b>Background:</b> Cutaneous leishmaniasis (CL) is a prevalent infectious zoonotic disease between humans and animals. Golestan Province is one of the important centers of CL in Iran. The current method for detecting <i>Leishmania</i> parasites in patients is Giemsa-stained direct smear from skin ulcers; however, PCR is a strongly recommended method for epidemiological studies. This study aimed to investigate <i>Leishmania</i> species in negative ulcer smears from patients suspected of having CL referred to Aq Qala Health Center using the multiplex nested PCR method.<b><span style="font-size:9.0pt"></span></b><br> <b>Methods:</b> This study was performed on 72 negative ulcer smears from patients suspected of having CL referred to Aq Qala Health Center, Golestan Province, northeastern Iran, from August 2019 to April 2020, using the multiplex nested PCR method to detect <i>Leishmania major</i> and <i>Leishmania tropica</i> species.<b><span style="font-size:9.0pt"></span></b><br> <b>Results:</b> Out of 72 samples, 4 (5.55%) samples were positive by multiplex nested PCR. Moreover, all positive samples are related to <i>Leishmania major </i>species.<br> <b>Conclusion</b>: The multiplex nested PCR method is strongly recommended for detecting Leishmania species in negative ulcer smears from patients suspected of having CL.</span></span><br> &nbsp; Leishmaniasis, Cutaneous, Leishmania major, Leishmania tropica, Negative Ulcer Smear, Multiplex Nested PCR 6 8 http://mlj.goums.ac.ir/browse.php?a_code=A-10-1375-2&slc_lang=en&sid=1 Fatemeh Livani dr_flivani@yahoo.com 100319475328460029989 100319475328460029989 No Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran Faramarz Koohsar fkoohsar@gmail.com 100319475328460029990 100319475328460029990 No Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran Farideh Tohidi articleinformation2@gmail.com 100319475328460029991 100319475328460029991 No Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran Mitra Sharbatkhori allauthore@gmail.com 100319475328460029992 100319475328460029992 No Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran Roghiyeh Faridnia r_faridnia400@yahoo.com 100319475328460029993 100319475328460029993 No Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran Ganesh Yadagiri 100319475328460029994 100319475328460029994 No Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA Ayeneh Hajieh Pangh ayenehpangh56@gmail.com 100319475328460029995 100319475328460029995 No Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran Mehdi Khoshrou articlesubmitt@yahoo.com 100319475328460029996 100319475328460029996 No Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran Hamed Kalani hamed.kalani@yahoo.com 100319475328460029997 100319475328460029997 Yes Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran