Phát hiện ô nhiễm mycoplasma bằng phản ứng chuỗi polymerase

Cytotechnology - Tập 16 - Trang 67-77 - 1994
Manfred Wirth1, Evelyn Berthold2, Martina Grashoff1, Horst Pfützner2, Uli Schubert3, Hansjörg Hauser1
1Genetik von Eukaryonten, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Germany
2Institute for Veterinary Medicine (Robert von Ostertag Institute) Jena Branch, Federal Health Office, Jena, Germany
3Institute for Medical Immunology, Medical Division (Charité), Humboldt-University of Berlin, Berlin, Germany

Tóm tắt

Phản ứng chuỗi polymerase (PCR) đã được sử dụng để phát hiện chung các loại Mollicutes. 25 loài Mycoplasma và Acholeplasma đã được phát hiện, bao gồm các tác nhân ô nhiễm quan trọng trong nuôi cấy tế bào như M. orale, M. arginini, M. hyorhinis, M. fermentans, A. laidlawii và các mycoplasma của con người và động vật khác. Các phản ứng PCR được thực hiện bằng cách sử dụng một bộ mồi nested được xác định từ các vùng bảo tồn của gen 16S rRNA. Giới hạn phát hiện được xác định là 1 fg DNA mycoplasma, tương đương với 1–2 bản sao genome của vùng mã hóa 16S rRNA. Danh tính của các sản phẩm khuếch đại đã được xác nhận bằng phương pháp điện di gel agarose và phân tích enzyme cắt giới hạn. DNA từ các vi sinh vật có mối quan hệ gần gũi và xa lạ không tạo ra các sản phẩm khuếch đại cụ thể. Phương pháp được trình bày ở đây cung cấp một xét nghiệm nhạy cảm, cụ thể và nhanh chóng hơn nhiều để phát hiện mycoplasmas so với những phương pháp hiện có.

Từ khóa

#PCR #mycoplasma #phát hiện ô nhiễm #vi sinh vật.

Tài liệu tham khảo

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