کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7556985 | 1491302 | 2018 | 25 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Duplex recombinase polymerase amplification assays incorporating competitive internal controls for bacterial meningitis detection
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کلمات کلیدی
RFUBHIRPAHDAIACPOCTHFNTCDNA - DNA یا اسید دزوکسی ریبونوکلئیکdeoxyribonucleic acid - اسید deoxyribonucleicNo template control - بدون کنترل قالبTetrahydrofuran - تتراهیدروفورانbrain heart infusion - تزریق قلب مغزیLoop-mediated isothermal amplification - تقویت ایزوترمال متصل به حلقهRecombinase polymerase amplification - تقویت پلیمراز RecombinaseLOD یا Limit of detection - حد تشخیصDuplex - دوبلکسLAMP - لامپCSF - مایع مغزی نخاعیCerebrospinal fluid - مایع مغزی نخاعیlimit of detection - محدودیت تشخیصBacterial meningitis - مننژیت باکتریاییpoint-of-care - نقطه مراقبتRelative Fluorescence Units - واحد فلورسانس نسبیpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازPirB - پیربhigh performance liquid chromatography - کروماتوگرافی مایع با کارایی بالاHPLC - کروماتوگرافی مایعی کاراInternal amplification control - کنترل تقویت داخلی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
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چکیده انگلیسی
Recombinase polymerase amplification (RPA) is an isothermal nucleic acid amplification technology that provides rapid and robust infectious disease pathogen detection, ideal for point-of-care (POC) diagnostics in disease-prevalent low-resource countries. We have developed and evaluated three duplex RPA assays incorporating competitive internal controls for the detection of leading bacterial meningitis pathogens. Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae singleplex RPA assays were initially developed and evaluated, demonstrating 100% specificity with limits of detection of 4.1, 8.5 and 3.9 genome copies per reaction, respectively. Each assay was further developed into internally controlled duplex RPA assays via the incorporation of internal amplification control templates. Clinical performance of each internally controlled duplex RPA assay was evaluated by testing 64 archived PCR-positive clinical samples. Compared to real-time PCR, all duplex RPA assays demonstrated 100% diagnostic specificity, with diagnostic sensitivities of 100%, 86.3% and 100% for the S. pneumoniae, N. meningitidis and H. influenzae assays, respectively. This study details the first report of internally controlled duplex RPA assays for the detection of bacterial meningitis pathogens: S. pneumoniae, N. meningitidis and H. influenzae. We have successfully demonstrated the clinical diagnostic utility of each duplex RPA assay, introducing effective diagnostic technology for POC bacterial meningitis identification in disease-prevalent developing countries.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 546, 1 April 2018, Pages 10-16
Journal: Analytical Biochemistry - Volume 546, 1 April 2018, Pages 10-16
نویسندگان
Owen Higgins, Eoin Clancy, Matthew S. Forrest, Olaf Piepenburg, Martin Cormican, Teck Wee Boo, Nicola O'Sullivan, Claire McGuinness, Deirdre Cafferty, Robert Cunney, Terry J. Smith,