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Reverse-Transcription Loop-Mediated Isothermal Amplification Has High Accuracy for Detecting Severe Acute Respiratory Syndrome Coronavirus 2 in Saliva and Nasopharyngeal/Oropharyngeal Swabs from Asymptomatic and Symptomatic Individuals.

J Mol Diagn

April 2022

Hampshire Hospitals National Health Service (NHS) Foundation Trust, Department of Microbiology, Basingstoke and North Hants Hospital, Basingstoke, United Kingdom; NHS Test and Trace Programme, Department of Health and Social Care, London, United Kingdom.

Previous studies have described reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for the rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal/oropharyngeal swab and saliva samples. This multisite clinical evaluation describes the validation of an improved sample preparation method for extraction-free RT-LAMP and reports clinical performance of four RT-LAMP assay formats for SARS-CoV-2 detection. Direct RT-LAMP was performed on 559 swabs and 86,760 saliva samples and RNA RT-LAMP on extracted RNA from 12,619 swabs and 12,521 saliva samples from asymptomatic and symptomatic individuals across health care and community settings.

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Article Synopsis
  • Researchers wanted to find a fast way to test for COVID-19 (SARS-CoV-2) among people who may or may not show symptoms.
  • They tested healthcare workers every week using a new method called LamPORE, which combines two types of technologies: loop-mediated amplification and nanopore sequencing.
  • The results showed that LamPORE was very accurate, catching nearly all cases in symptomatic people (100% sensitivity) and almost all in asymptomatic people (over 99.5% sensitivity).
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We describe the optimisation of a simplified sample preparation method which permits rapid and direct detection of SARS-CoV-2 RNA within saliva, using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Treatment of saliva samples prior to RT-LAMP by dilution 1:1 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98 °C heat step for 2 min enabled detection of SARS-CoV-2 RNA in positive saliva samples. Using RT-LAMP, SARS-CoV-2 RNA was detected in as little as 05:43 min, with no amplification detected in 3097 real-time reverse transcription PCR (rRT-PCR) negative saliva samples from staff tested within a service evaluation study, or for other respiratory pathogens tested (n = 22).

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Fine needle aspiration cytology (FNAC) is an important tool in the investigation of thyroid nodules and has few reported complications. We present the first report of recurrent laryngeal nerve palsy arising as a complication of thyroid nodule FNAC. This complication led to inaccurate diagnosis and unnecessarily radical surgery, with consequent increased morbidity.

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