Development of a pan-rickettsial molecular diagnostic test based on recombinase polymerase amplification assay

Kissenkötter, Jonas; Hansen, Søren Gustenhoff; Böhlken-Fascher, Susanne; Ademowo, Olusegun G.; Oyinloye, Oladapo Elijah; Bakarey, Adeleye Solomon; Dobler, Gerhard; Tappe, Dennis; Patel, Pranav; Czerny, Claus-Peter; Wahed, Ahmed Abd El

doi:10.1016/j.ab.2017.12.018

Cited by 20 publications

(9 citation statements)

References 21 publications

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“…RPA has the ability to tolerate mismatches, and the highest mismatch tolerability reported so far is nine nucleotide base pairs across the primer and probe binding sites. [82][83][84][85][86][87][88][89] Studies also showed that the mismatches at the 5′-end or centre of primers only mildly affect the RPA reaction, but mismatches located at the 3′-end of primers significantly affect the reaction. 84,86 This is consistent with the RPA reaction mechanism (see section 2.2), since the polymerase extends the primers and probe (once cleaved) from the 3′-terminus.…”

Section: Tolerability To Mismatches Inhibitors and Background Dnamentioning

confidence: 99%

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Macdonald

Stetten

2019

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480

365

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Section: Tolerability To Mismatches Inhibitors and Background Dnamentioning

confidence: 99%

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Macdonald

Stetten

2019

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480

365

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“…DNA was extracted from all sera samples and tested for 16S rRNA gene of Rickettsia spp. by real-time PCR ( 11 ). Sera samples were tested by IFA (Vircell, Spain) for detection of antibodies against R. conorii .…”

Section: Case Historymentioning

confidence: 99%

Cases of Mediterranean spotted fever in southeast of Iran

Farrokhnia¹,

Ghalejoogh²,

Rohani³

et al. 2020

IJM

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In this study the clinical manifestations, laboratory findings, and management of five patients diagnosed with Mediterra- nean spotted fever (MSF) from southeast of Iran are presented. All patients but one had recent tick-bite histories which were noticeable as black eschars (tache noire). Patients’ samples were tested by real-time PCR and serology (IFA). The disease was confirmed by fourfold rising of IgG antibodies against Rickettsia conorii. This is the first report of MSF cases in Iran.

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“…Since RPA was reported for the first time in 2006, it has been widely applied to the detection of viruses (Abd El Wahed et al 2015;Tu et al 2017;Gao et al 2020;Miao et al 2019), parasites (Sun et al 2016;Kersting et al 2014), and foodborne pathogens (Gao et al 2016;Kissenkötter et al 2018;Geng et al 2018;Hu et al 2019); identification of genetically modified crops (Xu et al 2014;Liu et al 2020);and medical diagnosis (Euler et al 2012;Boyle et al 2013). In comparison, the application of RPA to detect and identify harmful algae is in its infancy (Table 1).…”

Section: Introductionmentioning

confidence: 99%

Development of a recombinase polymerase amplification combined with lateral flow dipstick assay for rapid and sensitive detection of Heterosigma akashiwo

et al. 2021

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Heterosigma akashiwo is one of the main toxin-producing species that is globally known for frequently forming fish-killing harmful algal blooms. In this study, a novel technique referred to as recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) (RPA-LFD) was established for detection of H. akashiwo. RPA primers and an LFD probe were designed based on the sequence of the large ribosomal subunit (LSU rDNA) D1-D2 region of H. akashiwo. RAP/RPA-LFD was experimentally verified to be specific, displaying no cross-reaction with other control microalgae. It was demonstrated that the optimal amplification temperature and time for RPA were 41 °C and 40 min. The LFD assay was completed in only 5-10 min to analyze RPA products, and RPA results could be visualized directly. RPA-LFD was 100 times more sensitive than PCR, displaying a detection limit of 3.37 × 10 −4 ng µL −1 for genomic DNA of H. akashiwo and 1.28 × 10 2 copies µL −1 for the recombinant plasmid containing the inserted LSU rDNA D1-D2 region of H. akashiwo. In this study, RPA-LFD's detection limit was 1 cell mL −1 , which is feasible to apply in the field test. Thus, the established RPA-LFD targeting the detection of H. akashiwo is characterized by simplicity, rapidity, sensitivity, specificity, and visualization.

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Development of a pan-rickettsial molecular diagnostic test based on recombinase polymerase amplification assay

Cited by 20 publications

References 21 publications

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Cases of Mediterranean spotted fever in southeast of Iran

Development of a recombinase polymerase amplification combined with lateral flow dipstick assay for rapid and sensitive detection of Heterosigma akashiwo

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