Evaluation of sample treatments in a safe and straightforward procedure for the detection of SARS-CoV-2 in saliva

Rubio, Camila Peres; Franco‐Martínez, Lorena; Resalt, Cristina Sánchez; Torres-Cantero, Alberto M.; Martínez-Morata, Irene; Bernal, Enrique; Alcaraz, María José; Vicente-Romero, María Rosario; Martı́nez-Subiela, Silvia; Cerón, José J.

doi:10.1016/j.ijid.2021.05.053

Cited by 7 publications

(9 citation statements)

References 24 publications

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“…This method has also been modified replacing the use of proteinase K with a temperature gradient by heating at 95˚C/75˚C [31]. Saliva swabs have been reported having high sensitivity and specificity for the detection of influenza virus by the Xpert Xpress Flu/RSV test with a high overall agreement and Ct correlation with nasopharyngeal specimens [32]. In the present work, saliva samples were treated with proteinase K. A preliminary LoD for each assay was determined testing in triplicate (n = 3) samples of RNA purified using each extraction method, SalivaDirect TM [18] and in house Laboratory Developed Test (LDT).…”

Section: Resultsmentioning

confidence: 99%

Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples

Martínez¹,

Nunley²,

Gaines³

et al. 2022

JBM

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To contain the rapid and global spread of SARS-CoV-2, it is essential to develop an accurate and sensitive test system to address pandemic bottlenecks, simplified sample collection, and no sample prep. While meeting the demand of testing large populations, the miniaturized volume of assay reagents and offering rapid results is the need in such scenarios. Moreover, in view of the reports of co-infections and overlapping symptoms of influenza caused by Influenza A or Influenza B, and COVID-19 caused by SARS-CoV-2, a test system with three targets can be supportive for accurate clinical diagnosis. In this presentation, we evaluated the performance of a test comprising Microchip RT-PCR Influenza and COVID-19 Detection System for identifying these three viral pathogens in nasal swabs and saliva specimens. A rapid and simplified total nucleic acid extraction method was developed and validated for the reliable, high-throughput simultaneous detection of respiratory viruses causing Influenza (type A and type B viruses) and COVID-19 (SARS-CoV-2 virus) using the microchip-based AriaDNA TM platform deriving the name ABC Test. The test system was evaluated using 81 nasal swab samples, 77 clinical saliva samples, 5 blind CAP reference samples, and RNA standards. The limit of detection (LoD) was assessed using SARS-CoV-2, Influenza A, and Influenza B RNA standards. The multiplex ABC Test microchip displayed LoD of 14 copies/µL for SARS-CoV-2 and approximately 26 copies/µL for influenza A, and 140 copies/µL for influenza B, respectively. The ABC Test offers rapid multiplex one-step RT-PCR in 32 minutes for 45 cycles as the miniaturized reaction of 1.2 µL offering a highly sensitive, robust, and accurate assay for the detection of influenza A/B, and SARS-CoV-2.

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Section: Resultsmentioning

confidence: 99%

Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples

Martínez¹,

Nunley²,

Gaines³

et al. 2022

JBM

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“…Detergent treatments for potentially infectious matrices were introduced into the manufacturing processes of medical products more than 20 years ago [ 23 ] and have contributed to the overall safety of laboratory procedures. Additionally, when compared with heat inactivation procedures, detergents seem to have less significant impacts on the clinical chemistry and the analysis of hematology parameters [ 24 ]. Although NP-40 has been used less extensively for virus inactivation, Triton X-100 is a well-established method because of its lack of interference in the analysis of biofluids, such as human plasma and blood [ 25 ].…”

Section: Discussionmentioning

confidence: 99%

“…Although NP-40 has been used less extensively for virus inactivation, Triton X-100 is a well-established method because of its lack of interference in the analysis of biofluids, such as human plasma and blood [ 25 ]. Hersberger et al, [ 26 ] additionally showed that the latter does not seem to interfere with the performance of most chemical and hematological assays; though, Rubio and Franco-Martínez [ 24 ] reported that its presence at 0.5% concentration might interfere with the colorimetric lamp. However, the possible influence of the detergents used in this study on the analysis of porcine serum biomarkers and/or analytical tests remains to be studied.…”

Section: Discussionmentioning

confidence: 99%

Impact of ASFV Detergent Inactivation on Biomarkers in Serum and Saliva Samples

et al. 2022

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African swine fever (ASF) is a notifiable viral disease of domestic and wild suids. Despite intensive research efforts, the pathogenesis of the disease is still far from being understood. Analysis of biomarkers in different body fluids may supplement traditional pathogenesis studies. As reliable protocols are often established in laboratories with lower biosafety, the reliable inactivation of samples is crucial. The objective of this study was to find a procedure that inactivates the virus while preserving the biomarkers for downstream analyses. To this means, three different inactivation protocols were employed, namely Tergitol-type NP-40 (NP-40), polyoxyethylene-p-t-octylphenol (Triton X-100) and one with 95 °C heating. It could be demonstrated that all samples treated with 0.5% (v/v) concentration of both detergents showed an absence of virus infectivity. The same was true for heated samples. However, heated serum was not suitable for analyses. Next, the impact of treatment on biomarker readouts was assessed. While all protocols had an impact on the detection of biomarkers, correlation was retained. In particular, NP-40 may be the desired detergent for more accurate measurements while achieving efficient virus inactivation. Based on these studies, samples can be reliably inactivated for most biomarker analyses, and thus broader interdisciplinary cooperation is possible.

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“…Saliva has gained special importance in recent years as a diagnostic fluid since it is easy to obtain by a non-invasive sampling process. The SARS-CoV-2 virus can be detected in this fluid as well as the immunoglobulins directly produced after the immune response 3 – 5 . Overall, saliva could serve as a possible source of potential biomarkers in the clinical management of patients with COVID-19 5 , 6 .…”

Section: Introductionmentioning

confidence: 99%

“…The SARS-CoV-2 virus can be detected in this fluid as well as the immunoglobulins directly produced after the immune response 3 – 5 . Overall, saliva could serve as a possible source of potential biomarkers in the clinical management of patients with COVID-19 5 , 6 .…”

Section: Introductionmentioning

confidence: 99%

Saliva changes in composition associated to COVID-19: a preliminary study

Muñoz-Prieto

Rubić

González-Sánchez

et al. 2022

Sci Rep

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The coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV 2), is usually associated with a wide variety of clinical presentations from asymptomatic to severe cases. The use of saliva as a diagnostic and monitoring fluid has gained importance since it can be used to investigate the immune response and to direct quantification of antibodies against COVID-19. Additionally, the use of proteomics in saliva has allowed to increase our understanding of the underlying pathophysiology of diseases, bringing new perspectives on diagnostics, monitoring, and treatment. In this work, we compared the salivary proteome of 10 patients with COVID-19, (five patients with mild and five patients with severe COVID-19) and ten control healthy patients. Through the application of proteomics, we have identified 30 proteins whose abundance levels differed between the COVID-19 groups and the control group. Two of these proteins (TGM3 and carbonic anhydrase-CA6) were validated by the measurement of gGT and TEA respectively, in 98 additional saliva samples separated into two groups: (1) COVID-19 group, integrated by 66 patients who tested positive for COVID-19 (2) control group, composed of 32 healthy individuals who did not show any sign of disease for at least four weeks and were negative for COVID-19 in RT-PCR. In the proteomic study there were observed upregulations in CAZA1, ACTN4, and ANXA4, which are proteins related to the protective response against the virus disturbance, and the upregulation of TGM3, that is correlated to the oxidative damage in pulmonary tissue. We also showed the downregulation in cystatins and CA6 that can be involved in the sensory response to stimulus and possibly related to the presence of anosmia and dysgeusia during the COVID-19. Additionally, the presence of FGB in patients with severe COVID-19 but not in mild COVID-19 patients could indicate a higher viral aggregation and activation in these cases. In conclusion, the salivary proteome in patients with COVID-19 showed changes in proteins related to the protective response to viral infection, and the altered sensory taste perception that occur during the disease. Moreover, gGT and TEA could be potential biomarkers of respiratory complications that can occurs during COVID 19 although further larger studies should be made to corroborate this.

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Evaluation of sample treatments in a safe and straightforward procedure for the detection of SARS-CoV-2 in saliva

Cited by 7 publications

References 24 publications

Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples

Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples

Impact of ASFV Detergent Inactivation on Biomarkers in Serum and Saliva Samples

Saliva changes in composition associated to COVID-19: a preliminary study

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Evaluation of sample treatments in a safe and straightforward procedure for the detection of SARS-CoV-2 in saliva

Cited by 7 publications

References 24 publications

Validation of Microchip Based RT-PCR ABC Test (InfA/B &amp; COVID-19) in Clinical Samples

Validation of Microchip Based RT-PCR ABC Test (InfA/B &amp; COVID-19) in Clinical Samples

Impact of ASFV Detergent Inactivation on Biomarkers in Serum and Saliva Samples

Saliva changes in composition associated to COVID-19: a preliminary study

Contact Info

Product

Resources

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Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples

Validation of Microchip Based RT-PCR ABC Test (InfA/B & COVID-19) in Clinical Samples