Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Common Strains of
            <i>Escherichia coli</i>

Hill, Joshua; Beriwal, Shilpa; Ishwad, Chandra; Paul, Vinod K.; Kapil, Anish; Singh, Tripti; Wadowsky, Robert M.; Singh, Vinita; Goyal, Ankur; Jahnukainen, Timo; Johnson, James R.; Tarr, Phillip I.; Vats, Abhay

doi:10.1128/jcm.00152-08

Cited by 237 publications

(208 citation statements)

References 22 publications

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“…A melt-curve was run as a control to differentiate the product from nonspecific products, such as primer dimers. 45 …”

Section: Pcr In Labreadermentioning

confidence: 99%

“…To assemble a device, two mirror-image enclosure units are snapped together and placed over the circuit board containing the LEDs and detectors ( Figure S7). T he optical 45 setup and electronics are aligned in the cover by springs inside the enclosure, which force the parts against reference components. In this study, temperature control units, as well as automated data analysis methods were developed and integrated into the LabReader, in order to enable automated DNA 50 amplification, readout and analysis (see SI).…”

Section: Dna Purification In the Labtubementioning

confidence: 99%

“…19 , 20 T hey further often require expensive hardware for optical readout or fluidic control and are usable for one kind of application only. 15,19 In this paper, we introduce the LabSystem, a semi-automated 45 and frugal DNA p urification, amplification and readout testing system for diagnostic and quality control applications (Figure 1). The LabSystem consists of the LabT ube, a disposable cartridge for automation of DNA purification inside laboratory centrifuges, and the LabReader, a low-cost, handheld UV/Vis reader, to 50 amplify, readout and analyze the purified DNA.…”

Section: Introductionmentioning

confidence: 99%

“…40 Unlike PCR, LAMP does not require thermal cycling [41][42][43] , allowing a simpler and cheaper, disposable heating system to be used , [44][45][46][47][48] , and it is particularly temperature robust 40,48,49 . We 80 observed the LAMP reaction to be stable at temperatures of 65±5°C (T able S-6).…”

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confidence: 99%

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A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

Hoehl

Bocholt

Kloke³

et al. 2014

Analyst

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Contamination of foods is a public health hazard that episodically causes thousands of deaths and sickens millions worldwide. T o ensure food safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification, 10 amplification and detection. It consists of a disposable, centrifugally -driven DNA purification platform (LabT ube) and the subsequent amplification and detection in a low-cost UV/vis-reader (LabReader). For demonstration of the LabSystem in the context of food safety, purification of Escherichia coli (nonpathogenic E. coli and pathogenic verotoxin-producing E. coli (VT EC)) in water and milk, and the product-spoiler Alicyclobacillus acidoterrestris (A. acidoterrestris) in apple juice was integrated and 15 optimized in the LabT ube. Inside the LabReader, the purified DNA was amplified, readout and analyzed using both qualitative isothermal loop-mediated DNA amplification (LAMP) and quantitative real-time PCR. For the LAMP -LabSystem, the combined detection limits for purification and amplification of externally lysed VT EC and A. acidoterrestris is 10 2 -10 3 cell-equivalents. In the PCR-LabSystem for E. coli cells, the quantification limit is 10 2 cell-equivalents including LabT ube-integrated lysis. T he 20 demonstrated LabSystem only requires a laboratory centrifuge (to operate the disposable, fully closed LabT ube) and the low-cost LabReader for DNA amplification, readout and analysis. Compared with commercial DNA amplification devices, the LabReader improves sensitivity and specificity by the simultaneous readout of four wavelengths and the continuous readout during temperature cycling. T he use of a detachable eluate tube as an interface affords semi-automation of the LabSystem, which does not 25 require specialized training. It reduces hands-on time from about 50 to 3 min with only two handling steps: sample input and transfer of the detachable detection tube.

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“…A melt-curve was run as a control to differentiate the product from nonspecific products, such as primer dimers. 45 …”

Section: Pcr In Labreadermentioning

confidence: 99%

Section: Dna Purification In the Labtubementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

Hoehl

Bocholt

Kloke³

et al. 2014

Analyst

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“…As a first example, VT EC DNA amplification was integrated into the LabT ube. T he LAMP reaction (HaraKudo et al 2007) was chosen over other amplification methods, such as PCR (Hill et al 2008). This is because the desired test result does not have to be quantitative (a simple yes or no answer above a certain threshold suffices (Europäische Kommission 2004, Gill and Ghaemi 2 008)), yet it has to be sensitive, temperature robust and specific for the target organism.…”

Section: Heating Requirementsmentioning

confidence: 99%

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Hoehl

Weißert

Dannenberg

et al. 2014

Biomed Microdevices

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Abstract:This paper introduces a disposable battery-driven heating system for loop-mediated isothermal DNA amplification (LAMP) inside a centrifugally-driven DNA p urification platform (LabT ube). We demonstrate LabT ube-based fully automated DNA purification of as low as 100 cell-equivalents of verotoxin-producing Escherichia coli (VT EC) in water, milk and apple juice in a laboratory centrifuge, followed by integrated and automated LAMP amplification with a reduction of hands-on time from 45 to 1min. The heating system consists of two parallel SMD thick film resistors and a NT C as heating and temperature sensing elements. They are driven by a 3V battery and controlled by a microcontroller. The LAMP reagents are stored in the elution chamber and the amplification starts immediately after the eluate is purged into the chamber. The LabT ube, including a microcontroller-based heating system, demonstrates contamination-free and automated sample-to-answer nucleic acid testing within a laboratory centrifuge. The heating system can be easily parallelized within one LabT ube and it is deployable for a variety of heating and electrical applications.

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Culture‐free biphasic approach for sensitive detection of Escherichia coli O157:H7 from beef samples

Mostafa

Ganguli

Berger

et al. 2021

Biotech & Bioengineering

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Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Common Strains of Escherichia coli

Cited by 237 publications

References 22 publications

A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Culture‐free biphasic approach for sensitive detection of Escherichia coli O157:H7 from beef samples

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