Đánh giá kháng nguyên tái tổ hợp hemolysin co-regulated protein 1 (hcp1) trong chẩn đoán nhanh bệnh nhân nhiễm melioidosis (bệnh Whitmore) bằng kỹ thuật ELISA

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Tác giả: Anh Đao Tran, Anh Tram Que, Tinh Trinh Ho, Hai Linh Bui Nguyen, Trung Hoang Quang, Quoc Tuan Do, Trung Trinh Thanh, Huyen Pham Thi, Le Quyen Tran Thi, Tinh Nguyen Thi

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học và Công nghệ Việt Nam - B, 2020

Mô tả vật lý: tr.26

Bộ sưu tập: Metadata

ID: 309160

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Melioidosis (also called Whitmore’s disease) is a fatal infectious disease caused by a Gram-negative soil-dwelling bacterium Burkholderia pseudomallei. Due to the diverse clinical manifestations, final diagnosis of the disease mostly relies on the examination of bacterial culture. However, the sensitivity of the culture method depends on various factors, leading to the misdiagnosis of the disease in the clinical setting. In this study, the authors amplified BPSS1498 gene coding hemolysin co-regulated protein 1 (hcp1), cloned and expressed the recombinant protein using E. coli BL21(DE3) pLysS. The recombinant hcp1 protein antigen with 18.7 kDa was then purified and demonstrated a highly specific binding with human IgG of the culture-confirmed patient with melioidosis using Western blot. Hcp1-ELISA was established and evaluated on 94 sera collected from 32 culture-confirmed patients with melioidosis, 32 patients infected with other bacteria, and 30 healthy persons. Using a cut-off value of 0.595, hcp1-ELISA showed a sensitivity of 93.7% and a specificity of 98.4%. Those results lead to a potential application of hcp1-ELISA in the diagnosis of melioidosis in Vietnam, especially on the diagnosis of the disease in patients with fever of unknown origin due to the limitation of the routine culture methods.Melioidosis (hay còn gọi là bệnh Whitmore) là một loại bệnh truyền nhiễm nguy hiểm do vi khuẩn Burkholderia pseudomallei (B. pseudomallei) gây nên. Bệnh có biểu hiện lâm sàng đa dạng. Vì vậy, chẩn đoán khẳng định ca nhiễm bệnh phải dựa trên bằng chứng xét nghiệm vi sinh lâm sàng. Tuy nhiên, độ nhạy của phương pháp xét nghiệm nuôi cấy thường phụ thuộc vào nhiều yếu tố, dẫn đến nguy cơ bỏ sót ca bệnh trong quá trình xét nghiệm. Trong nghiên cứu này, các tác giả xác định gene BPSS1498 mã hóa hemolysin co-regulated protein 1 (hcp1), tạo dòng và biểu hiện protein bằng tế bào E. coli BL21(DE3) pLysS. Protein hcp1 có kích thước 18,7 kDa được tinh sạch. Western blot chứng minh hcp1 gắn kết đặc hiệu với kháng thể IgG của bệnh nhân nhiễm melioidosis. Phương pháp hcp1-ELISA được thiết lập và đánh giá trên 94 mẫu huyết thanh (32 mẫu từ nhóm bệnh nhân melioidosis, 32 mẫu từ nhóm bệnh nhân nhiễm khuẩn huyết khác và 30 mẫu từ nhóm người khỏe mạnh). Sử dụng giá trị cut-off là 0,595, kỹ thuật hcp1-ELISA đạt độ nhạy 93,7% và độ đặc hiệu 98,4%. Kết quả nghiên cứu này mở ra hướng chẩn đoán mới trong xét nghiệm sàng lọc bệnh nhân nhiễm melioidosis tại Việt Nam, trên đối tượng bệnh nhân sốt kéo dài chưa rõ nguyên nhân do những hạn chế khi nuôi cấy vi khuẩn B. pseudomallei từ mẫu bệnh phẩm lâm sàng.

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