Isothermal Amplification Technology for Disease Diagnosis

Boonbanjong, Poramin; Treerattrakoon, Kiatnida; Waiwinya, Wassa; Pitikultham, Piyawat; Japrung, Deanpen

doi:10.3390/bios12090677

Cited by 30 publications

(15 citation statements)

References 60 publications

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“…Several isothermal amplification methods have been developed for identifying infectious agents including rolling circle amplification (RCA), recombinase polymerase amplification (RPA), multiple displacement amplification (MDA), ladder-shape melting temperature isothermal amplification, and helicase-dependent amplification (HDA). These diagnostic methods have been used in the fields of agriculture, medicine, and veterinary medicine (as reviewed by Boonbanjong et al 16 ). Among these, LAMP is one of the most widely used methods.…”

Section: Discussionmentioning

confidence: 99%

Detection of Babesia bovis using loop-mediated isothermal amplification (LAMP) with improved thermostability, sensitivity and alternative visualization methods

Arnuphapprasert

Nugraheni

Aung

et al. 2023

Sci Rep

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Bovine babesiosis is one of the most economically important tick-borne diseases in tropical and subtropical countries. A conventional microscopic diagnosis is typically used because it is inexpensive and expeditious. However, it is highly dependent on well-trained microscopists and tends to be incapable of detecting subpatent and chronic infections. Here, we developed a novel nucleic acid-based amplification method using loop-mediated isothermal amplification (LAMP) in conjunction with a colori-fluorometric dual indicator for the rapid and accurate detection of Babesia bovis based on the mitochondrial cytochrome b gene. We aimed to improve the thermostability, sensitivity, specificity, and alternative visualization of LAMP-based methods. We assessed its diagnostic performance compared to two conventional PCR agarose gel electrophoresis (PCR-AGE) methods. The thermostability of LAMP reaction mixtures and DNA templates in variable conditions was also assessed. In addition, we evaluated alternative visualization methods using different light sources including neon, LED, and UV lights. We found that the LAMP-neon was ten times more sensitive than the PCR-AGE, while the LAMP-LED and LAMP-UV were 1,000 times more sensitive. The current LAMP method showed no cross-amplification with uninfected cattle DNA or other common blood parasites in cattle, including Babesia bigemina, Theileria orientalis, Anaplasma marginale, and Trypanosoma evansi. In addition, the developed LAMP method has good thermostability and the potential for on-site utility as B. bovis DNA could still be detected up to 72 h after initial preparation. Our findings suggested that the developed LAMP method provides an alternative approach for B. bovis detection with sensitivity higher than PCR-AGE diagnostics, high specificity, and the flexibility to use neon, LED, and UV light sources for positive signal observations.

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Section: Discussionmentioning

confidence: 99%

Detection of Babesia bovis using loop-mediated isothermal amplification (LAMP) with improved thermostability, sensitivity and alternative visualization methods

Arnuphapprasert

Nugraheni

Aung

et al. 2023

Sci Rep

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“…A novel anti-NS2BNS3pro antibody-based indirect enzyme-linked immunosorbent assay (ELISA) showed good sensitivity (87%) and reasonable specificity (76%) ( 11 ). Diagnostics based on the CRISPR/Cas13 system allow the fast, highly sensitive, and specific detection of DENV ( 12 – 15 ). Among new technologies for clinical diagnostics, reverse transcription-recombinase polymerase amplification (RT-RPA) does not require thermal cycling instrumentation for nucleic acid amplification, showing the potential to provide rapid, low-resource detection that can be adapted for serotype-specific detection ( 16 – 19 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices

et al. 2023

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“…DNA amplification using RCA is cost-effective, simple, and at the same time can be highly specific . RCA has shown great potential for use in diagnostics and as a biosensor, and various biologically relevant targets have been detected using RCA. − Hence, it is not surprising that isothermal systems such as RCA are anticipated to be used for on site testing in remote areas and poorly equipped healthcare settings . Besides being used in biosensing, RCA is currently one of the most promising DNA replication methods for the construction of a synthetic cell. − To that end, however, a major drawback is that the output DNA structure of a RCA reaction is different to the input structure, that is linear instead of circular DNA.…”

Section: Introductionmentioning

confidence: 99%

C2CAplus: A One-Pot Isothermal Circle-to-Circle DNA Amplification System

Grasemann,

Thiel Pizarro,

Maerkl

2023

ACS Synth. Biol.

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Rolling circle amplification (RCA) is a widely used DNA amplification method that uses circular template DNA as input and produces multimeric, linear single-or double-stranded DNA. Circle-to-circle amplification (C2CA) has further expanded this method by implementing product recircularization using restriction and ligation, leading to a higher amplification yield and enabling the generation of circular products. However, C2CA is a multistep, nonisothermal method, requiring multiple fluid manipulations and thereby compromises several advantages of RCA.Here, we improved C2CA to implement a one-pot, single step, isothermal reaction at temperatures ranging from 25 to 37 °C. Our C2CAplus method is simple, robust, and produces large quantities of product DNA that can be seen with the naked eye.

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Isothermal Amplification Technology for Disease Diagnosis

Cited by 30 publications

References 60 publications

Detection of Babesia bovis using loop-mediated isothermal amplification (LAMP) with improved thermostability, sensitivity and alternative visualization methods

Detection of Babesia bovis using loop-mediated isothermal amplification (LAMP) with improved thermostability, sensitivity and alternative visualization methods

Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices

C2CAplus: A One-Pot Isothermal Circle-to-Circle DNA Amplification System

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