RPB0544

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
N. meningitidis Neisseria meningitidis,personal::Sara E. Branham M1027,"Diplokokkus intracellularis meningitidis","Micrococcus meningitidis","Neisseria weichselbaumii","Micrococcus intracellularis","Micrococcus meningitidis cerebrospinalis" 487 Neisseriales Neisseriaceae Neisseria Neisseria meningitidis Bacteria

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM) GC Content(%) Predicted Melting Temperature(℃) Molecular Weight(g/moles) Positions in GenBank accession number
F GTC AGG ATA AAT GGA TTG CTC AAG GTT A 28 \ 39.29 56.36 8691.73 \
R CGC ATT CGA CAC ATA CAA TAC ATC TTT A 28 \ 35.71 54.63 8475.61 \

Gene Description

Target Gene GenBank ID
ctrA gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
Our approach combined open-source modular automation (Opentrons) with adaptable molecular biology protocols, enabling a versatile, rapid, and reliable diagnostic workflow for infectious diseases. RPA \ 20 min 37 °C \ \ \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 An Automated Versatile Diagnostic Workflow for Infectious Disease Detection in Low-Resource Settings Miren Urrutia Iturritza,Phuthumani Mlotshwa,Jesper Gantelius,Tobias Alfvén,Edmund Loh,Jens Karlsson,Chris Hadjineophytou,Krzysztof Langer,Konstantinos Mitsakakis,Aman Russom,Håkan N Jönsson,Giulia Gaudenzi Micromachines 38930678 10.3390/mi15060708

An Automated Versatile Diagnostic Workflow for Infectious Disease Detection in Low-Resource Settings

Author(s):

Miren Urrutia Iturritza,Phuthumani Mlotshwa,Jesper Gantelius,Tobias Alfvén,Edmund Loh,Jens Karlsson,Chris Hadjineophytou,Krzysztof Langer,Konstantinos Mitsakakis,Aman Russom,Håkan N Jönsson,Giulia Gaudenzi

Journal:

Micromachines

Year:

2024

Abstract:

Laboratory automation effectively increases the throughput in sample analysis, reduces human errors in sample processing, as well as simplifies and accelerates the overall logistics. Automating diagnostic testing workflows in peripheral laboratories and also in near-patient settings -like hospitals, clinics and epidemic control checkpoints- is advantageous for the simultaneous processing of multiple samples to provide rapid results to patients, minimize the possibility of contamination or error during sample handling or transport, and increase efficiency. However, most automation platforms are expensive and are not easily adaptable to new protocols. Here, we address the need for a versatile, easy-to-use, rapid and reliable diagnostic testing workflow by combining open-source modular automation (Opentrons) and automation-compatible molecular biology protocols, easily adaptable to a workflow for infectious diseases diagnosis by detection on paper-based diagnostics. We demonstrated the feasibility of automation of the method with a low-cost Neisseria meningitidis diagnostic test that utilizes magnetic beads for pathogen DNA isolation, isothermal amplification, and detection on a paper-based microarray. In summary, we integrated open-source modular automation with adaptable molecular biology protocols, which was also faster and cheaper to perform in an automated than in a manual way. This enables a versatile diagnostic workflow for infectious diseases and we demonstrated this through a low-cost N. meningitidis test on paper-based microarrays.