Clinical Applications of the Polymerase Chain Reaction: An Update

Cuchacovich, Raquel

doi:10.1016/j.idc.2006.09.003

Cited by 12 publications

(8 citation statements)

References 97 publications

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“…Since the introduction of molecular diagnostics and in particular real-time PCR, ample proof of its sensitivity and specificity has been generated. Indeed, molecular diagnostics are deemed superior to bacterial culture techniques or serological diagnostics (2)(3)(4). It has even been suggested to entirely eliminate the old methods in order to streamline centralized laboratories for molecular diagnostics (5)(6)(7).…”

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confidence: 99%

Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

et al. 2013

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Syndromic panels for infectious disease have been suggested to be of value in point-of-care diagnostics for developing countries and for biodefense. To test the performance of isothermal recombinase polymerase amplification (RPA) assays, we developed a panel of 10 RPAs for biothreat agents. The panel included RPAs for Francisella tularensis, Yersinia pestis, Bacillus anthracis, variola virus, and reverse transcriptase RPA (RT-RPA) assays for Rift Valley fever virus, Ebola virus, Sudan virus, and Marburg virus. Their analytical sensitivities ranged from 16 to 21 molecules detected (probit analysis) for the majority of RPA and RT-RPA assays. A magnetic bead-based total nucleic acid extraction method was combined with the RPAs and tested using inactivated whole organisms spiked into plasma. The RPA showed comparable sensitivities to real-time RCR assays in these extracts. The run times of the assays at 42°C ranged from 6 to 10 min, and they showed no cross-detection of any of the target genomes of the panel nor of the human genome. The RPAs therefore seem suitable for the implementation of syndromic panels onto microfluidic platforms. Syndromic panels for infectious and emerging infectious diseases have been suggested to be of value in point-of-care (POC) diagnostics for developing countries and for biodefense (1). Since the introduction of molecular diagnostics and in particular real-time PCR, ample proof of its sensitivity and specificity has been generated. Indeed, molecular diagnostics are deemed superior to bacterial culture techniques or serological diagnostics (2-4). It has even been suggested to entirely eliminate the old methods in order to streamline centralized laboratories for molecular diagnostics (5-7).In recent years alternative isothermal amplification methods which can be categorized into (i) T7 promoter-driven amplifications (transcription-mediated amplification [TMA], nucleic acid sequence-based amplification [NASBA], and single primer isothermal amplification [SPIA]), (ii) strand displacement methods (strand displacement amplification [SDA], loop-mediated isothermal amplification [LAMP], and smart amplification [SmartAmp]), (iii) helicase-dependent amplification (HDA), (iv) recombinase polymerase amplification (RPA), and (v) rolling-circle amplification (RCA) methods (8-12) have been developed. Some were purposely designed for isothermal amplification starting from RNA (TMA, NASBA, and SPIA), whereas others initially targeted DNA (SDA, LAMP, HDA, RPA, and RCA) and were only later adapted for RNA targets. Nonspecific intercalating fluorophores or fluorescent primers have been used for real-time detection in LAMP, SDA, HDA, and RCA, and specific detection probe formats have been developed for NASBA, RCA, HDA,.In isothermal and exponential RPA, the phage recombinase UvsX and its cofactor UvsY form a nucleoprotein complex with oligonucleotide primers to scan for homologous sequences in a DNA template. Recognition of a specific homologous sequence leads to the initiation of strand invasion of the com...

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confidence: 99%

Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

et al. 2013

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“…Indeed partly molecular diagnostics are deemed superior to bacterial culture techniques or serological diagnostics 16,18,19 . Some authors have even opted for entirely eliminating the old methods to streamline centralised laboratories for molecular diagnostics 1,20,21 .…”

Section: Minaturising Molecular Diagnostic Assaysmentioning

confidence: 99%

“…Each province is divided into urban and rural communes. At the communal level, five rural communes share the Sourou Valley: Barani (47,991 inhabitants) and Sono (7,317) on the western bank; Di (23,864), Lanfièra (18,817) and Gassan (37,873) …”

Section: Study Areamentioning

confidence: 99%

“…Over 50% of individuals examined had urinary schistosomiasis, with majority being males aged > 5 years old [17]. The distribution of the disease in the region is mainly determined by the distributions of the intermediate snail hosts [18,19]. Schistosoma mansoni is highly endemic immediately along the southern shore of the lake and decreases with distance from the shorelines [15].…”

Section: Epidemiology Of Schistosomiasis Around the Lake Victoriamentioning

confidence: 99%

“…Poor nutritional status, an immunocomprised state and malignancies are factors contributing to the poor outcome. The clinical incidence varies from region to region, but has been reported up to 2.2% of the hospital admissions 18 .…”

Section: Pyogenic Liver Abscessmentioning

confidence: 99%

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