Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
Por:
Miranda-Ortiz, Haydee, Fernandez-Figueroa, Edith A., Ruiz-Garcia, Erika B., Munoz-Rivas, Anallely, Mendez-Perez, Alejandra, Mendez-Galvan, Jorge, de la Vega, Horacio Astudillo, Gabino-Lopez, Belem, Nava-Monroy, Ricardo, la Torre, Alejandro Lopez-Flores a, Lopez-Vergara Anaya, Tanit, Vilar-Compte, Diana, Coquis-Navarrete, Uriel, Valdes-Reyes, Melissa, Sanchez-Montes, Sokani, Becker, Ingeborg
Publicada:
1 ene 2021
Categoría:
Multidisciplinary
Resumen:
The use of saliva for the diagnosis of SARS-CoV-2 has shown to be a good
alternative to nasopharyngeal swabs (NPS), since it permits
self-collection, avoids the exposure of healthy persons to infected
patients, reduces waiting times, eliminates the need of personal
protective equipment and is non-invasive. Yet current saliva testing is
still expensive due to the need of specialized tubes containing buffers
to stabilize the RNA of SARS-CoV-2 and inactivate the virus. These tubes
are expensive and not always accessible in sufficient quantities. We now
developed an alternative saliva testing method, using TRIzol for
extraction, viral inactivation, and storage of SARS-CoV-2 RNA, combined
with RT-qPCR, which was comparable in its performance to NPS. Paired
saliva samples and NPS were taken from 15 asymptomatic healthcare
workers and one patient with SARS-CoV-2. Further 13 patients with
SARS-CoV-2 were only saliva-tested. All the tests were performed
according to CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR
Diagnostic Panel. Saliva (4 mL) was taken in sterile 50 mL tubes, 1.5 mL
TRIzol were added and mixed. Our results show that 5 mu L of saliva RNA
extracted with TRIzol allow for an adequate detection of the virus in
patients positive for SARS-CoV-2 and was equally sensitive to NPS in
TRIzol. We conclude that saliva testing using TRIzol is a recommendable
method for diagnosis of SARS-CoV-2 since it has several advantages over
currently used saliva tests: it can be done with normal sterile tubes,
does not need cold-chain handling, is stable at room temperature, is
non-invasive and less costly, making it more accessible for low-income
countries. Cheaper saliva testing using TRIzol is especially relevant
for low-income countries to optimize diagnosis and help define
quarantine durations for families, healthcare workers, schools, and
other public workplaces, thus decreasing infections and mortality caused
by SARS-CoV-2.
Filiaciones:
Miranda-Ortiz, Haydee:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Munoz-Rivas, Anallely:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Mendez-Perez, Alejandra:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Mendez-Galvan, Jorge:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
de la Vega, Horacio Astudillo:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Gabino-Lopez, Belem:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Nava-Monroy, Ricardo:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
la Torre, Alejandro Lopez-Flores a:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Lopez-Vergara Anaya, Tanit:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Vilar-Compte, Diana:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Coquis-Navarrete, Uriel:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Valdes-Reyes, Melissa:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Sanchez-Montes, Sokani:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
Becker, Ingeborg:
Fernandez-Figueroa, EA (Corresponding Author), Natl Inst Genom Med, Computat & Integrat Gen, Mexico City, DF, Mexico. Fernandez-Figueroa, EA
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