A high throughput immuno-affinity mass spectrometry method for detection and quantitation of SARS-CoV-2 nucleoprotein in human saliva and its comparison with RT-PCR, RT-LAMP, and lateral flow rapid antigen test.

Autor:	Lane D; The Department of Chemical Pathology and Metabolic Diseases, Leicester Royal Infirmary, University Hospitals of Leicester, Leicester, UK.; Department of Cardiovascular Sciences, University of Leicester, Leicester, UK., Allsopp R; Department of Genetics and Genome Biology, Leicester Cancer Research Centre, University of Leicester, Leicester, UK., Holmes CW; Clinical Microbiology, Leicester Royal Infirmary, University Hospitals of Leicester NHS Trust, Leicester, UK., Slingsby OC; van Geest MS-OMICS Facility, University of Leicester, Leicester, UK., Jukes-Jones R; The Department of Chemical Pathology and Metabolic Diseases, Leicester Royal Infirmary, University Hospitals of Leicester, Leicester, UK., Bird P; Clinical Microbiology, Leicester Royal Infirmary, University Hospitals of Leicester NHS Trust, Leicester, UK., Anderson NL; SISCAPA Assay Technologies, Inc., Washington, DC, USA., Razavi M; SISCAPA Assay Technologies, Inc., Washington, DC, USA., Yip R; SISCAPA Assay Technologies, Inc., Washington, DC, USA., Pearson TW; SISCAPA Assay Technologies, Inc., Washington, DC, USA., Pope M; SISCAPA Assay Technologies, Inc., Washington, DC, USA., Khunti K; Leicester Diabetes Centre, Leicester General Hospital, University of Leicester, Leicester, UK., Doykov I; Genetics & Genomic Medicine Department, Translational Mass Spectrometry Research Group, UCL Institute of Child Health, London, UK.; Great Ormond Street Biomedical Research Centre, UCL Institute of Child Health, London, UK., Hällqvist J; Genetics & Genomic Medicine Department, Translational Mass Spectrometry Research Group, UCL Institute of Child Health, London, UK.; Great Ormond Street Biomedical Research Centre, UCL Institute of Child Health, London, UK., Mills K; Genetics & Genomic Medicine Department, Translational Mass Spectrometry Research Group, UCL Institute of Child Health, London, UK.; Great Ormond Street Biomedical Research Centre, UCL Institute of Child Health, London, UK., Skipp P; Centre for Proteomic Research, University of Southampton, Southampton, UK., Carling R; Biochemical Sciences, Synnovis, Guys & St Thomas' NHSFT, London, UK.; GKT School Medical Education, Kings College London, London, UK., Ng L; Department of Cardiovascular Sciences, University of Leicester, Leicester, UK.; van Geest MS-OMICS Facility, University of Leicester, Leicester, UK., Shaw J; Department of Genetics and Genome Biology, Leicester Cancer Research Centre, University of Leicester, Leicester, UK., Gupta P; The Department of Chemical Pathology and Metabolic Diseases, Leicester Royal Infirmary, University Hospitals of Leicester, Leicester, UK.; Department of Cardiovascular Sciences, University of Leicester, Leicester, UK., Jones DJL; Department of Genetics and Genome Biology, Leicester Cancer Research Centre, University of Leicester, Leicester, UK.; van Geest MS-OMICS Facility, University of Leicester, Leicester, UK.
Jazyk:	angličtina
Zdroj:	Clinical chemistry and laboratory medicine [Clin Chem Lab Med] 2024 Jan 23; Vol. 62 (6), pp. 1206-1216. Date of Electronic Publication: 2024 Jan 23 (Print Publication: 2024).
DOI:	10.1515/cclm-2023-0243
Abstrakt:	Objectives: Many reverse transcription polymerase chain reaction (RT-PCR) methods exist that can detect SARS-CoV-2 RNA in different matrices. RT-PCR is highly sensitive, although viral RNA may be detected long after active infection has taken place. SARS-CoV-2 proteins have shorter detection windows hence their detection might be more meaningful. Given salivary droplets represent a main source of transmission, we explored the detection of viral RNA and protein using four different detection platforms including SISCAPA peptide immunoaffinity liquid chromatography-mass spectrometry (SISCAPA-LC-MS) using polyclonal capture antibodies. Methods: The SISCAPA-LC MS method was compared to RT-PCR, RT-loop-mediated isothermal amplification (RT-LAMP), and a lateral flow rapid antigen test (RAT) for the detection of virus material in the drool saliva of 102 patients hospitalised after infection with SARS-CoV-2. Cycle thresholds (Ct) of RT-PCR ( E gene) were compared to RT-LAMP time-to-positive (TTP) ( NE and Orf1a genes), RAT optical densitometry measurements (test line/control line ratio) and to SISCAPA-LC-MS for measurements of viral protein. Results: SISCAPA-LC-MS showed low sensitivity (37.7 %) but high specificity (89.8 %). RAT showed lower sensitivity (24.5 %) and high specificity (100 %). RT-LAMP had high sensitivity (83.0 %) and specificity (100.0 %). At high initial viral RNA loads (<20 Ct), results obtained using SISCAPA-LC-MS correlated with RT-PCR (R 2 0.57, p-value 0.002). Conclusions: Detection of SARS-CoV-2 nucleoprotein in saliva was less frequent than the detection of viral RNA. The SISCAPA-LC-MS method allowed processing of multiple samples in <150 min and was scalable, enabling high throughput. (© 2023 Walter de Gruyter GmbH, Berlin/Boston.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu