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2022, Cilt 52, Sayı 1, Sayfa(lar) 073-081 |
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Development of Dual and Single PCR Protocols for Intestinal Helminths: A Preliminary Study on Multiplex PCR |
Hayriye Kırkoyun Uysal1, Sinem Öktem Okullu2, Elif Merve Aydın3, Eray Şahin4, Bahar Akgün Karapınar1, Özgür Kurt2 |
1İstanbul Üniversitesi, Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, İstanbul, Türkiye 2Acıbadem Mehmet Ali Aydınlar Üniversitesi, Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, İstanbul, Türkiye 3Koç Üniversitesi, Sağlık Bilimleri Enstitüsü, Hücresel ve Moleküler Tıp Doktora Programı, İstanbul, Türkiye 4Acıbadem Mehmet Ali Aydınlar Üniversitesi, Sağlık Bilimleri Enstitüsü, Biyoistatistik ve Biyoinformatik Programı, İstanbul, Türkiye |
Keywords: Turkey, Intestinal helminth, Multiplex PCR, Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis, Taenia spp., Strongyloides stercoralis |
Objective: The aim of this initial study of a project is to identify some intestinal helminths which have limited
diagnostic options despite their known associations with fatal and chronic clinical cases in the world and in our
country, using multiplex PCR.
Method: Using the DNAs of Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis, Taenia spp. and Strongyloides stercoralis which have been received from a foreign center for joint studies, single and double optimization trials of the helminths were made within the project. Results: In the end of the study period, single optimization of T. trichiura (280 bp) as well as double optimizations of A. lumbricoides (995 bp) - S. stercoralis (115 bp) and Taenia spp. (950 bp) - E. vermicularis (400 bp) were completed successfully, and their diagnostic protocols were described. Conclusion: Two different duplex, and a single PCR protocol have been completed in the initial study. The aim in the next term will be both to increase the number of helminths within the PCR protocol, and determination of the sensitivity and specificity of the tests using the stool samples of healthy persons and patients in new studies. These efforts will hopefully contribute to better understanding the epidemiology and roles of intestinal helminths in clinical cases. |
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