Smartphone-Based
in-Gel Loop-Mediated Isothermal Amplification
(gLAMP) System Enables Rapid Coliphage MS2 Quantification in Environmental
Waters
Version 2 2018-05-16, 13:56
Version 1 2018-05-16, 13:48
Posted on 2018-05-16 - 13:56
Model
coliphages (e.g., ΦX174, MS2, and PRD1) have been widely
used as surrogates to study the fate and transport of pathogenic
viruses in the environment and during wastewater treatment. Two
groups of coliphages (F-specific and somatic) are being explored as
indicators of viral fecal pollution in ambient water. However, the
detection and quantification of coliphages still largely rely on time-consuming
culture-based plaque assays. In this study, we developed an in-gel
loop-mediated isothermal amplification (gLAMP) system enabling coliphage
MS2 quantification within 30 min using standard laboratory devices.
Viral particles (MS2) were immobilized with LAMP reagents in polyethylene
glycol hydrogel, and then viral RNAs were amplified through a LAMP
reaction. Due to the restriction effect of the hydrogel matrix, one
viral particle would only produce one amplicon dot. Therefore, the
sample virus concentrations can be determined based on the number
of fluorescent amplicon dots using a smartphone for imaging. The method
was validated by using artificially spiked and naturally contaminated
water samples. gLAMP results were shown to correlate well with plaque
assay counts (R2 = 0.984, p < 0.05) and achieved similar sensitivity to quantitative reverse-transcription
polymerase chain reaction (RT-qPCR; 1 plaque-forming unit per reaction).
Moreover, gLAMP demonstrated a high level of tolerance against inhibitors
naturally present in wastewater, in which RT-qPCR was completely inhibited.
Besides MS2, gLAMP can also be used for the quantification of other
microbial targets (e.g., Escherichia coli and Salmonella). Considering its simplicity, sensitivity, rapidity,
and versatility, gLAMP holds great potential for microbial water-quality
analysis, especially in resource-limited settings.
CITE THIS COLLECTION
DataCite
3 Biotech
3D Printing in Medicine
3D Research
3D-Printed Materials and Systems
4OR
AAPG Bulletin
AAPS Open
AAPS PharmSciTech
Abhandlungen aus dem Mathematischen Seminar der Universität Hamburg
ABI Technik (German)
Academic Medicine
Academic Pediatrics
Academic Psychiatry
Academic Questions
Academy of Management Discoveries
Academy of Management Journal
Academy of Management Learning and Education
Academy of Management Perspectives
Academy of Management Proceedings
Academy of Management Review
Huang, Xiao; Lin, Xingyu; Urmann, Katharina; Li, Lijie; Xie, Xing; Jiang, Sunny; et al. (2018). Smartphone-Based
in-Gel Loop-Mediated Isothermal Amplification
(gLAMP) System Enables Rapid Coliphage MS2 Quantification in Environmental
Waters. ACS Publications. Collection. https://doi.org/10.1021/acs.est.8b00241
or
Select your citation style and then place your mouse over the citation text to select it.
SHARE
Usage metrics
AUTHORS (7)
XH
Xiao Huang
XL
Xingyu Lin
KU
Katharina Urmann
LL
Lijie Li
XX
Xing Xie
SJ
Sunny Jiang
MH
Michael R. Hoffmann
KEYWORDS
amplicon dotculture-based plaque assayswater-quality analysisPRDpathogenic virusesEnvironmental Waters Model coliphagesin-gel loop-mediated. TwoR 2gLAMP resultsSystem Enables Rapid Coliphage MS 2 Quantificationambient waterRNAsample virus concentrationsrestriction effectplaque assay countsresource-limited settingslaboratory devicesLAMP reagents1 plaque-forming unitamplicon dotswastewater treatment . Two groupsLAMP reactionViral particlespolyethylene glycol hydrogelSmartphone-Based in-Gel Loop-Mediated Isothermal Amplification30 minreverse-transcription polymerase chain reactionhydrogel matrixwater samplescoliphage MS 2 quantification