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Detection of sub-mi...
Detection of sub-microscopic blood levels of Plasmodium falciparum using Tandem Oligonucleotide Repeat Cascade Amplification (TORCA) assay with an attomolar detection limit
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- Ghindilis, Andrey L. (författare)
- TORCATECH LLC, 5210,104th St SW, Mukilteo, WA 98275 USA
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- Chesnokov, Olga (författare)
- Florida Atlantic Univ, Charles E Schmidt Coll Med, Dept Biomed Sci, 777 Glades Rd, Boca Raton, FL 33428 USA
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- Ngasala, Billy (författare)
- Uppsala universitet,Internationell barnhälsa och nutrition,Muhimbili Univ Hlth & Allied Sci, Dept Parasitol & Med Entomol, Dar Es Salaam, Tanzania
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- Smith, Maria W. (författare)
- TORCATECH LLC, 5210,104th St SW, Mukilteo, WA 98275 USA
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- Smith, Kenneth (författare)
- TORCATECH LLC, 5210,104th St SW, Mukilteo, WA 98275 USA
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- Mårtensson, Andreas, 1963- (författare)
- Uppsala universitet,Internationell barnhälsa och nutrition
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- Oleinikov, Andrew V. (författare)
- Florida Atlantic Univ, Charles E Schmidt Coll Med, Dept Biomed Sci, 777 Glades Rd, Boca Raton, FL 33428 USA
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(creator_code:org_t)
- 2019-02-27
- 2019
- Engelska.
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Ingår i: Scientific Reports. - : NATURE PUBLISHING GROUP. - 2045-2322. ; 9
- Relaterad länk:
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https://doi.org/10.1...
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https://uu.diva-port... (primary) (Raw object)
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https://www.nature.c...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Tandem Oligonucleotide Repeat Cascade Amplification (TORCA) based on signal rather than target amplification under isothermal conditions was developed for nucleic acid assays. The initial signal was generated by hybridization of single stranded DNA targets to immobilized recognition probes followed by hybrid cleavage with specific restriction endonuclease (REase), and release of trigger oligonucleotides (Tr1). The signal amplification chamber contained two bead types carrying single-stranded amplification probes and two amplification REases. The probes consisted of multiple tandem repeats of either Tr1 or another trigger Tr2, with the tandem-Tr1 anchored to the beads through the antisense Tr2 linker and vice versa. Addition of the recognition reaction solution and Tr1 hybridization to the anti-Tr1 linkers started cleavage and release of additional Tr1 and Tr2, resulting in exponential signal amplification. The cleavage cascade also released horseradish peroxidase (HRP) pre-attached to the amplification probes, and the resultant signal was measured colorimetrically. A TORCA assay was developed for detection of Plasmodium falciparum parasites in blood. It had the detection limit in the attomolar concentration range, successfully detecting sub-microscopic P. falciparum infections at less than 0.75 infected erythrocytes per microliter. Further TORCA optimization will likely produce the quantitative isothermal alternative to PCR at a fraction of its cost.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
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