WHotLAMP: A simple, inexpensive, and sensitive molecular test for the detection of SARS-CoV-2 in saliva
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Abstract
Despite the development of effective vaccines against SARS-CoV-2, epidemiological control of the virus is still challenging due to slow vaccine rollouts, incomplete vaccine protection to current and emerging variants, and unwillingness to get vaccinated. Therefore, frequent testing of individuals to identify early SARS-CoV-2 infections, contact-tracing and isolation strategies remain crucial to mitigate viral spread. Here, we describe WHotLAMP, a rapid molecular test to detect SARS-CoV-2 in saliva. WHotLAMP is simple to use, highly sensitive (~4 viral particles per microliter of saliva) and specific, as well as inexpensive, making it ideal for frequent screening. Moreover, WHotLAMP does not require toxic chemicals or specialized equipment and thus can be performed in point-of-care settings, and may also be adapted for resource-limited environments or home use. While applied here to SARS-CoV-2, WHotLAMP can be modified to detect other pathogens, making it adaptable for other diagnostic assays, including for use in future outbreaks.
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SciScore for 10.1101/2021.06.17.21259050: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: Collection and processing of patient nasal and saliva samples: With informed consent (Columbia University IRB AAAT1974), patients were provided sterile cotton tipped swabs and conical tubes for sample collection. Sex as a biological variable not detected. Randomization not detected. Blinding To test the sensitivity of WHotLAMP, patient saliva samples and negative control samples were tested under blind conditions. Power Analysis not detected. Table 2: Resources
Software and Algorithms Sentences Resources Saliva RNA spike-in assay: For saliva RNA spike-in tests, a 1.7 mL tube with 100 μL of saliva was combined with 100 μL lysis buffer (0.8 M guanidine hydrochloride (G3272, Sigma), 2% Tween-20 … SciScore for 10.1101/2021.06.17.21259050: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: Collection and processing of patient nasal and saliva samples: With informed consent (Columbia University IRB AAAT1974), patients were provided sterile cotton tipped swabs and conical tubes for sample collection. Sex as a biological variable not detected. Randomization not detected. Blinding To test the sensitivity of WHotLAMP, patient saliva samples and negative control samples were tested under blind conditions. Power Analysis not detected. Table 2: Resources
Software and Algorithms Sentences Resources Saliva RNA spike-in assay: For saliva RNA spike-in tests, a 1.7 mL tube with 100 μL of saliva was combined with 100 μL lysis buffer (0.8 M guanidine hydrochloride (G3272, Sigma), 2% Tween-20 (BP337, Fisher Biotech)), mixed, and incubated at room temperature for 5 min. Fisher Biotech)suggested: (Resource Identification Portal, RRID:SCR_004098)For each sample, 100 μL of saliva was loaded into the tube, followed by 50 μL of RNAlater (R0901, Sigma) and 25 μL of Proteinase K (10 mg/mL) (PB0451, BioBasic; or 25530-049, Ambion), and mixed. BioBasicsuggested: NoneResults from OddPub: Thank you for sharing your code.
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a protocol registration statement.
Results from scite Reference Check: We found no unreliable references.
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