TY - JOUR

T1 - A Novel Multiplex LAMP Assay for the Detection of Respiratory Human Adenoviruses

AU - Koryukov, Maksim A.

AU - Oscorbin, Igor P.

AU - Novikova, Lidiya M.

AU - Gordukova, Maria A.

AU - Turina, Irina E.

AU - Galeeva, Elena V.

AU - Kudlay, Dmitry A.

AU - Filipenko, Maxim L.

N1 - This work was supported by the Russian state-funded project for ICBFM SB RAS (grant number 121031300045-2).

PY - 2024/7

Y1 - 2024/7

N2 - Human adenoviruses (HAdVs) are common pathogens that are associated with a variety of diseases, including respiratory tract infections (RTIs). Without reliable, fast, and cost-effective detection methods for HAdVs, patients may be misdiagnosed and inappropriately treated. To address this problem, we have developed a multiplex loop-mediated isothermal amplification (LAMP) assay for the detection of the species Human adenovirus B (HAdV-B), Human adenovirus C (HAdV-C) and Human adenovirus E (HAdV-E) that cause RTIs. This multiplexing approach is based on the melting curve analysis of the amplicons with a specific melting temperature for each HAdV species. Without the need for typing of HAdVs, the LAMP results can be visually detected using colorimetric analysis. The assay reliably detects at least 375 copies of HAdV-B and -C and 750 copies of HAdV-E DNA per reaction in less than 35 min at 60 °C. The designed primers have no in silico cross-reactivity with other human respiratory pathogens. Validation on 331 nasal swab samples taken from patients with RTIs showed a 90–94% agreement rate with our in-house multiplex quantitative polymerase chain reaction (qPCR) method. Concordance between the quantitative and visual LAMP was 99%. The novel multiplexed LAMP could be an alternative to PCR for diagnostic purposes, saving personnel and equipment time, or could be used for point-of-care testing.

AB - Human adenoviruses (HAdVs) are common pathogens that are associated with a variety of diseases, including respiratory tract infections (RTIs). Without reliable, fast, and cost-effective detection methods for HAdVs, patients may be misdiagnosed and inappropriately treated. To address this problem, we have developed a multiplex loop-mediated isothermal amplification (LAMP) assay for the detection of the species Human adenovirus B (HAdV-B), Human adenovirus C (HAdV-C) and Human adenovirus E (HAdV-E) that cause RTIs. This multiplexing approach is based on the melting curve analysis of the amplicons with a specific melting temperature for each HAdV species. Without the need for typing of HAdVs, the LAMP results can be visually detected using colorimetric analysis. The assay reliably detects at least 375 copies of HAdV-B and -C and 750 copies of HAdV-E DNA per reaction in less than 35 min at 60 °C. The designed primers have no in silico cross-reactivity with other human respiratory pathogens. Validation on 331 nasal swab samples taken from patients with RTIs showed a 90–94% agreement rate with our in-house multiplex quantitative polymerase chain reaction (qPCR) method. Concordance between the quantitative and visual LAMP was 99%. The novel multiplexed LAMP could be an alternative to PCR for diagnostic purposes, saving personnel and equipment time, or could be used for point-of-care testing.

KW - HAdV

KW - LAMP

KW - PCR

KW - adenoviruses

KW - melting curves

KW - multiplex

KW - qPCR

KW - respiratory infections

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85198359269&origin=inward&txGid=c2a2eeb2075a31c5e90350fdae0cdc8e

UR - https://www.mendeley.com/catalogue/15eab073-5870-3268-900e-46b56b3f8320/

U2 - 10.3390/ijms25137215

DO - 10.3390/ijms25137215

M3 - Article

C2 - 39000322

VL - 25

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1661-6596

IS - 13

M1 - 7215

ER -