Thiết lập và đánh giá quy trình sàng lọc trước sinh không xâm lấn cho các bệnh đơn gen trội phổ biến

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Tác giả: Thị Trang Đào, Thị Thanh Thủy Đỗ, Phước Lộc Đoàn, Thị Kim Phượng Đoàn, Hồng Chương Dương, Thị Minh Thi Hà, Hồng Thịnh Lê, Thị Lan Anh Lương, Cảnh Chương Nguyễn, Vạn Thông Nguyễn, Nhật Thăng Trần, Vũ Uyên Trần, Nhựt Thư Hương Trịnh

Ngôn ngữ: vie

Ký hiệu phân loại: 571.85 Developmental genetics

Thông tin xuất bản: Tạp chí Y học Việt Nam (Tổng hội Y học Việt Nam), 2022

Mô tả vật lý: 99-104

Bộ sưu tập: Metadata

ID: 498927

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Establish and validate a non-invasive prenatal testing protocol for early detection of some common dominant monogenic diseases by determining the sensitivity and specificity of the test. Method: A total of 30 blood samples from singleton pregnant women over 9 weeks of gestation, together with paternal blood samples were collected. Cell-free DNA were extracted from plasma samples, barcoded, enriched and then analyzed for 30 targeted genes by the Nextseq 2000 next-generation sequencing system. (Illumina, USA). The variant calls from the cell free DNA were compared with variants detected on paternal and maternal genomic DNA (trios analysis) to calculate the analytical sensitivity and specificity of the test. Results: We identified 29 true positive variants and 8 false positive variants in 30 samples. In contrast, more than 3 million variants within the 30 targeted genes had been correctly determined as true negative (both parents were homozygous of reference alleles). One false negative variant was observed on paternal genomic DNA but not on the corresponding cfDNA sample. With these numbers, we therefore reported that the analytical sensitivity of our test is 96.7% and analytical specificity is >
99%. Conclusion: We established a non-invasive prenatal testing protocol to screen for multiple single gene disorders with high accuracy. This is an important step towards expanding the scope of current noninvasive prenatal testing to detect a wide spectrum of dominant monogenic diseases.Thiết lập và đánh giá qui trình trước sinh không xâm lấn cho các bệnh di truyền trội đơn gen thông qua việc xác định độ nhạy và độ đặc hiệu kỹ thuật của xét nghiệm. Phương pháp: 30 mẫu máu ngoại vi của các thai phụ mang thai đơn trên 9 tuần thai kèm mẫu máu cha được thu nhận. DNA ngoại bào được tách chiết từ mẫu huyết tương, sau đó tiến hành tạo thư viện và lai-bắt giữ 30 gen mục tiêu và giải trình tự bằng hệ thống giải trình tự thế hệ mới Nextseq 2000 (Illumina, Hoa Kỳ). Các biến thể phát hiện trên DNA ngoại bào được so sánh với các biến thể phát hiện trên DNA nội bào của cha và mẹ (phân tích trios) để tính toán độ nhạy và độ đặc hiệu kỹ thuật của qui trình. Kết quả: Nghiên cứu phát hiện 29 biến thể dương tính thật và 8 biến thể dương tính giả trong tổng số 30 mẫu. Các biến thể dương tính giả xảy ra khi cả cha và mẹ đều có nucleotit chuẩn nhưng DNA ngoại bào cho thấy biến thể khác. Ngược lại, có hơn 3 triệu biến thể âm tính thật (cha mẹ đều đồng hợp tử nucleotit chuẩn) trong 30 gen mục tiêu đã được phát hiện chính xác. Có một biến thể âm tính giả được xác định khi tìm thấy trên mẫu DNA bộ gen của cha nhưng không phát hiện trên mẫu DNA ngoại bào tương ứng. Sử dụng thông số trên, nghiên cứu xác định độ nhạy kỹ thuật là 96.7% và độ đặc hiệu kỹ thuật là >
99%. Kết luận: Nghiên cứu đã thiết lập được qui trình sàng lọc trước sinh không xâm lấn bằng kỹ thuật giải trình tự thế hệ mới cho nhiều bệnh đơn gen với độ chính xác cao. Đây là tiền đề quan trọng tiến tới khả năng mở rộng phạm vi sàng lọc của xét nghiệm trước sinh không xâm lấn nhằm phát hiện các bệnh đơn gen trội phổ biến.

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