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1.
Microbiol Spectr ; 10(1): e0202921, 2022 02 23.
Article in English | MEDLINE | ID: covidwho-1673365

ABSTRACT

The objective of our study was to evaluate the sensitivity and specificity of rapid antigen detection tests versus those of reverse transcriptase PCR (RT-PCR) using oral, anterior nasal, and nasopharyngeal swabs. The underlying prospective, diagnostic case-control-type accuracy study included 87 hospitalized and nonhospitalized participants in a positive and a negative sample cohort between 16 March and 14 May 2021 in two hospitals in Vienna. SARS-CoV-2 infection status was confirmed by RT-PCR. Participants self-performed one oral and one anterior nasal swab for the rapid antigen test, immediately followed by two nasopharyngeal swabs for the rapid antigen test and RT-PCR by the investigator. Test results were read after 15 min, and participants completed a questionnaire in the meantime. Test parameters were calculated based on the evaluation of 87 participants. The overall sensitivity of rapid antigen detection tests versus that of RT-PCR with oral, anterior nasal, and nasopharyngeal samples was 18.18% (95% confidence interval [CI] 8.19% to 32.71%), 63.04% (95% CI 47.55% to 76.79%), and 73.33% (95% CI 58.06% to 85.4%), respectively. All sampling methods had a test specificity of 100% regardless of the cycle threshold (CT) value. Rapid antigen detection tests using self-collected anterior nasal swabs proved to be as sensitive as and more tolerable than professionally collected nasopharyngeal swabs for CT values up to 30 determined by RT-PCR. This finding illustrates the reliability of tests obtained by adequate self-collected anterior nasal specimen. Sensitivity was dependent upon the CT value for each sampling method. While the main advantage of rapid antigen detection tests is the immediate availability of results, PCR should be preferred in crucial settings wherever possible. IMPORTANCE Rapid antigen detection devices for SARS-CoV-2 represent a valuable tool for monitoring the spread of infection. However, the reliability of the tests depends largely on the test performance and the respective sampling method. Nasopharyngeal swabs mark the gold standard for sample collection in suspected respiratory tract infections but are unsuitable for widespread application, as they must be performed by medically trained personnel. With the underlying study, the head-to-head test performance and the usability of self-collected samples for SARS-CoV-2 detection using rapid antigen detection devices were evaluated. The results confirm similar sensitivity of self-collected anterior nasal swabs to that of professionally collected nasopharyngeal swabs for patients with a CT of < 30 determined by RT-PCR.


Subject(s)
COVID-19 Serological Testing/methods , COVID-19/diagnosis , Mouth/virology , Nasopharynx/virology , Nose/virology , SARS-CoV-2/isolation & purification , Adult , Aged , Aged, 80 and over , Antigens, Viral/analysis , COVID-19/virology , COVID-19 Nucleic Acid Testing , Female , Humans , Male , Middle Aged , Polymerase Chain Reaction , Prospective Studies , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Sensitivity and Specificity , Young Adult
2.
Int J Mol Sci ; 23(3)2022 Jan 27.
Article in English | MEDLINE | ID: covidwho-1667192

ABSTRACT

This review article was designed to evaluate the existing evidence related to the molecular processes of SARS-CoV-2 infection in the oral cavity. The World Health Organization stated that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and transmission is produced by respiratory droplets and aerosols from the oral cavity of infected patients. The oral cavity structures, keratinized and non-keratinized mucosa, and salivary glands' epithelia express SARS-CoV-2 entry and transmission factors, especially angiotensin converting enzyme Type 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2). Replication of the virus in cells leads to local and systemic infection spread, and cellular damage is associated with clinical signs and symptoms of the disease in the oral cavity. Saliva, both the cellular and acellular fractions, holds the virus particles and contributes to COVID-19 transmission. The review also presents information about the factors modifying SARS-CoV-2 infection potential and possible local pharmacotherapeutic interventions, which may confine SARS-CoV-2 virus entry and transmission in the oral cavity. The PubMed and Scopus databases were used to search for suitable keywords such as: SARS-CoV-2, COVID-19, oral virus infection, saliva, crevicular fluid, salivary gland, tongue, oral mucosa, periodontium, gingiva, dental pulp, ACE2, TMPRSS2, Furin, diagnosis, topical treatment, vaccine and related words in relevant publications up to 28 December 2021. Data extraction and quality evaluation of the articles were performed by two reviewers, and 63 articles were included in the final review.


Subject(s)
COVID-19/pathology , Mouth , SARS-CoV-2/physiology , Angiotensin-Converting Enzyme 2/metabolism , Angiotensin-Converting Enzyme 2/physiology , Animals , COVID-19/metabolism , COVID-19/transmission , COVID-19/virology , Humans , Mouth/metabolism , Mouth/pathology , Mouth/virology , Mouth Mucosa/metabolism , Mouth Mucosa/pathology , Mouth Mucosa/virology , Pathology, Oral , Receptors, Virus/genetics , Receptors, Virus/metabolism , SARS-CoV-2/pathogenicity , Serine Endopeptidases/metabolism , Serine Endopeptidases/physiology , Signal Transduction/genetics , Virus Internalization
3.
Viruses ; 14(1)2022 01 09.
Article in English | MEDLINE | ID: covidwho-1611142

ABSTRACT

We found and genetically described two novel SARS-like coronaviruses in feces and oral swabs of the greater (R. ferrumequinum) and the lesser (R. hipposideros) horseshoe bats in southern regions of Russia. The viruses, named Khosta-1 and Khosta-2, together with related viruses from Bulgaria and Kenya, form a separate phylogenetic lineage. We found evidence of recombination events in the evolutionary history of Khosta-1, which involved the acquisition of the structural proteins S, E, and M, as well as the nonstructural genes ORF3, ORF6, ORF7a, and ORF7b, from a virus that is related to the Kenyan isolate BtKY72. The examination of bats by RT-PCR revealed that 62.5% of the greater horseshoe bats in one of the caves were positive for Khosta-1 virus, while its overall prevalence was 14%. The prevalence of Khosta-2 was 1.75%. Our results show that SARS-like coronaviruses circulate in horseshoe bats in the region, and we provide new data on their genetic diversity.


Subject(s)
Chiroptera/virology , Severe acute respiratory syndrome-related coronavirus/genetics , Animals , Base Sequence , Chiroptera/classification , Evolution, Molecular , Feces/virology , Metagenomics , Mouth/virology , Phylogeny , Prevalence , Recombination, Genetic , Russia , Severe acute respiratory syndrome-related coronavirus/classification , Species Specificity , Spike Glycoprotein, Coronavirus/genetics , Viral Proteins/genetics
4.
EBioMedicine ; 73: 103675, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1596532

ABSTRACT

BACKGROUND: Following the discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its rapid spread throughout the world, new viral variants of concern (VOC) have emerged. There is a critical need to understand the impact of the emerging variants on host response and disease dynamics to facilitate the development of vaccines and therapeutics. METHODS: Syrian golden hamsters are the leading small animal model that recapitulates key aspects of severe coronavirus disease 2019 (COVID-19). We performed intranasal inoculation of SARS-CoV-2 into hamsters with the ancestral virus (nCoV-WA1-2020) or VOC first identified in the United Kingdom (B.1.1.7, alpha) and South Africa (B.1.351, beta) and analyzed viral loads and host responses. FINDINGS: Similar gross and histopathologic pulmonary lesions were observed after infection with all three variants. Although differences in viral genomic copy numbers were noted in the lungs and oral swabs of challenged animals, infectious titers in the lungs were comparable between the variants. Antibody neutralization capacities varied, dependent on the original challenge virus and cross-variant protective capacity. Transcriptional profiling of lung samples 4 days post-challenge (DPC) indicated significant induction of antiviral pathways in response to all three challenges with a more robust inflammatory signature in response to B.1.1.7 infection. Furthermore, no additional mutations in the spike protein were detected at 4 DPC. INTERPRETATIONS: Although disease severity and viral shedding were not significantly different, the emerging VOC induced distinct humoral responses and transcriptional profiles compared to the ancestral virus. These observations suggest potential differences in acute early responses or alterations in immune modulation by VOC. FUNDING: Intramural Research Program, NIAID, NIH; National Center for Research Resources, NIH; National Center for Advancing Translational Sciences, NIH.


Subject(s)
COVID-19/pathology , SARS-CoV-2/isolation & purification , Transcriptome , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , COVID-19/immunology , COVID-19/virology , Cricetinae , Dendritic Cells/cytology , Dendritic Cells/metabolism , Disease Models, Animal , Female , Immunity, Humoral , Lung/metabolism , Lung/pathology , Lung/virology , Mesocricetus , Mouth/pathology , Mouth/virology , Nucleocapsid Proteins/metabolism , RNA, Viral/analysis , RNA, Viral/metabolism , SARS-CoV-2/genetics , SARS-CoV-2/metabolism , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/immunology , Spike Glycoprotein, Coronavirus/metabolism
5.
Medicine (Baltimore) ; 100(51): e28327, 2021 Dec 23.
Article in English | MEDLINE | ID: covidwho-1592581

ABSTRACT

BACKGROUND: SARS-CoV-2 is the virus responsible for coronavirus disease-19 (COVID-19) disease, which has been shown to trigger multiple affectations. One of the first tissue areas to come into contact with the virus is the oral cavity, which develops various alterations. Hence, the objective of this systematic review was to identify the main signs and symptoms of this disease in the oral cavity, and the following research question was established: What are the main oral signs and symptoms in COVID-19-positive persons? METHODS: The electronic databases of PUBMED, SCOPUS, and SCIENCE DIRECT were analyzed, the keywords "ORAL DISEASES," "ORAL MANIFESTACTIONS," and "COVID-19" were used taking into account the following inclusion criteria: studies whose main objective was oral manifestations secondary to the confirmation of COVID-19, plus clinical cases, case series, and retrospective or prospective studies. For the assessment of the risk of bias the JBI Critical Appraisal Checklist for Case Series tool was used. RESULTS: A total of 18 studies were included, the most common initial signs/symptoms after contagion of SARS-CoV-2 were dysgeusia, dry mouth, and burning mouth, and the main signs/symptoms were the presence of ulcerative lesions, dysgeusia, and Candida albicans infections. CONCLUSIONS: It is very important to detect any alteration in the mucosa in patients with COVID-19 and to provide assertive treatment to avoid complications, and try to maintain adequate oral hygiene throughout the course of the disease to avoid the colonization of opportunistic microorganisms and to avoid complications both orally and systemically.


Subject(s)
COVID-19 , Mouth Diseases/virology , Mouth/virology , Candidiasis, Oral , Dysgeusia , Humans , Prospective Studies , Retrospective Studies
6.
J Med Microbiol ; 70(9)2021 Sep.
Article in English | MEDLINE | ID: covidwho-1393561

ABSTRACT

Introduction. Non-invasive sample collection and viral sterilizing buffers have independently enabled workflows for more widespread COVID-19 testing by reverse-transcriptase polymerase chain reaction (RT-PCR).Gap statement. The combined use of sterilizing buffers across non-invasive sample types to optimize sensitive, accessible, and biosafe sampling methods has not been directly and systematically compared.Aim. We aimed to evaluate diagnostic yield across different non-invasive samples with standard viral transport media (VTM) versus a sterilizing buffer eNAT- (Copan diagnostics Murrieta, CA) in a point-of-care diagnostic assay system.Methods. We prospectively collected 84 sets of nasal swabs, oral swabs, and saliva, from 52 COVID-19 RT-PCR-confirmed patients, and nasopharyngeal (NP) swabs from 37 patients. Nasal swabs, oral swabs, and saliva were placed in either VTM or eNAT, prior to testing with the Xpert Xpress SARS-CoV-2 (Xpert). The sensitivity of each sampling strategy was compared using a composite positive standard.Results. Swab specimens collected in eNAT showed an overall superior sensitivity compared to swabs in VTM (70 % vs 57 %, P=0.0022). Direct saliva 90.5 %, (95 % CI: 82 %, 95 %), followed by NP swabs in VTM and saliva in eNAT, was significantly more sensitive than nasal swabs in VTM (50 %, P<0.001) or eNAT (67.8 %, P=0.0012) and oral swabs in VTM (50 %, P<0.0001) or eNAT (58 %, P<0.0001). Saliva and use of eNAT buffer each increased detection of SARS-CoV-2 with the Xpert; however, no single sample matrix identified all positive cases.Conclusion. Saliva and eNAT sterilizing buffer can enhance safe and sensitive detection of COVID-19 using point-of-care GeneXpert instruments.


Subject(s)
COVID-19 Nucleic Acid Testing , Specimen Handling/methods , Adult , Aged , COVID-19/diagnosis , Containment of Biohazards , Culture Media , Female , Humans , Male , Middle Aged , Mouth/virology , Nasopharynx/virology , Nose/virology , Point-of-Care Testing , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Saliva/virology , Sensitivity and Specificity , Specimen Handling/standards
7.
Infection ; 49(3): 527-531, 2021 Jun.
Article in English | MEDLINE | ID: covidwho-1384713

ABSTRACT

OBJECTIVES: The optimal diagnostic specimen to detect SARS-CoV-2 by PCR in the upper respiratory tract is unclear. Mouthwash fluid has been reported as an alternative to nasopharyngeal and oropharyngeal swabs. We compared mouthwash fluid with a combined oro-nasopharyngeal swab regarding test performance. METHODS: In a large refugee facility, we retested individuals with a previous positive test for SARS-CoV-2 and their quarantined close contacts. All individuals were asymptomatic at the time of testing. First, a mouthwash (gargling for at least 5 s) with sterile water was performed. Then, with a single flocked swab the back of the throat and subsequently the nasopharynx were sampled. Samples were inactivated and analysed on a Roche cobas 6800® system with the Roche SARS-CoV-2 test. RESULTS: Of 76 individuals, 39 (51%) tested positive for SARS-CoV-2 by oro-nasopharyngeal swab. Mouthwash detected 13 of 76 (17%) infections, but did not detect any additional infection. Samples that were positive in both tests, had lower cycle threshold (Ct)-values for oro-nasopharyngeal samples, indicating a higher virus concentration, compared to samples only positive in oro-nasopharyngeal swabs. CONCLUSION: Mouthwash is not as sensitive as combined oro-nasopharyngeal swab in detecting upper respiratory tract infection.


Subject(s)
COVID-19 Nucleic Acid Testing , COVID-19/diagnosis , SARS-CoV-2/isolation & purification , Specimen Handling/methods , Adolescent , Adult , Asymptomatic Infections , Child , Female , Humans , Male , Middle Aged , Mouth/virology , Nasopharynx/virology , SARS-CoV-2/genetics , Sensitivity and Specificity , Young Adult
8.
J Med Virol ; 93(9): 5333-5338, 2021 Sep.
Article in English | MEDLINE | ID: covidwho-1363672

ABSTRACT

The accurate laboratory detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a crucial element in the fight against coronavirus disease 2019 (COVID-19). Reverse transcription-polymerase chain reaction testing on combined oral and nasopharyngeal swab (ONPS) suffers from several limitations, including the need for qualified personnel, the discomfort caused by invasive nasopharyngeal sample collection, and the possibility of swab and transport media shortage. Testing on saliva would represent an advancement. The aim of this study was to compare the concordance between saliva samples and ONPS for the detection of SARS-CoV-2 on various commercial and laboratory-developed tests (LDT). Individuals were recruited from eight institutions in Quebec, Canada, if they had SARS-CoV-2 RNA detected on a recently collected ONPS, and accepted to provide another ONPS, paired with saliva. Assays available in the different laboratories (Abbott RealTime SARS-CoV-2, Cobas® SARS-CoV-2, Simplexa™ COVID-19 Direct, Allplex™ 2019-nCoV, RIDA®GENE SARS-CoV-2, and an LDT preceded by three different extraction methods) were used to determine the concordance between saliva and ONPS results. Overall, 320 tests were run from a total of 125 saliva and ONPS sample pairs. All assays yielded similar sensitivity when saliva was compared to ONPS, with the exception of one LDT (67% vs. 93%). The mean difference in cycle threshold (∆C t ) was generally (but not significantly) in favor of the ONPS for all nucleic acid amplification tests. The maximum mean ∆​​​​​C t was 2.0, while individual ∆C t varied importantly from -17.5 to 12.4. Saliva seems to be associated with sensitivity similar to ONPS for the detection of SARS-CoV-2 by various assays.


Subject(s)
COVID-19 Nucleic Acid Testing/standards , COVID-19/diagnosis , Diagnostic Tests, Routine/standards , RNA, Viral/genetics , SARS-CoV-2/genetics , COVID-19/epidemiology , COVID-19/virology , COVID-19 Nucleic Acid Testing/instrumentation , COVID-19 Nucleic Acid Testing/methods , Diagnostic Tests, Routine/instrumentation , Diagnostic Tests, Routine/methods , Humans , Mouth/virology , Nasopharynx/virology , Quebec/epidemiology , Saliva/virology , Sensitivity and Specificity , Specimen Handling/standards
9.
Viruses ; 13(8)2021 07 30.
Article in English | MEDLINE | ID: covidwho-1335232

ABSTRACT

Since the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the susceptibility of animals and their potential to act as reservoirs or intermediate hosts for the virus has been of significant interest. Pigs are susceptible to multiple coronaviruses and have been used as an animal model for other human infectious diseases. Research groups have experimentally challenged swine with human SARS-CoV-2 isolates with results suggesting limited to no viral replication. For this study, a SARS-CoV-2 isolate obtained from a tiger which is identical to human SARS-CoV-2 isolates detected in New York City and contains the D614G S mutation was utilized for inoculation. Pigs were challenged via intravenous, intratracheal, or intranasal routes of inoculation (n = 4/route). No pigs developed clinical signs, but at least one pig in each group had one or more PCR positive nasal/oral swabs or rectal swabs after inoculation. All pigs in the intravenous group developed a transient neutralizing antibody titer, but only three other challenged pigs developed titers greater than 1:8. No gross or histologic changes were observed in tissue samples collected at necropsy. In addition, no PCR positive samples were positive by virus isolation. Inoculated animals were unable to transmit virus to naïve contact animals. The data from this experiment as well as from other laboratories supports that swine are not likely to play a role in the epidemiology and spread of SARS-CoV-2.


Subject(s)
COVID-19/virology , SARS-CoV-2/physiology , Administration, Intranasal , Administration, Intravenous , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , COVID-19/blood , COVID-19/immunology , Disease Models, Animal , Humans , Mouth/virology , Nose/virology , SARS-CoV-2/genetics , Swine , Trachea/virology , Virus Replication
10.
Virus Res ; 303: 198442, 2021 10 02.
Article in English | MEDLINE | ID: covidwho-1211167

ABSTRACT

OBJECTIVE: Association of SARS-CoV2 burden in the aerodigestive tract with the disease is sparsely understood. We propose to elucidate the implications of SARS-CoV2 copies in concurrent nasopharyngeal swab (NPS), whole mouth fluid (WMF) and respiratory droplet (RD) samples on disease pathogenesis/transmission. METHODS: SARS-CoV2 copies quantified by RT-PCR in concurrent NPS, WMF and RD samples from 80 suspected COVID-19 patients were analysed with demographics, immune response and disease severity. RESULTS: Among the 55/80 (69 %) NPS-positive patients, SARS-CoV2 was detected in 44/55 (80 %) WMF (concordance with NPS-84 %; p = 0.02) and 17/55 (31 %) RD samples. SARS-CoV2 copies were similar in NPS (median:8.74 × 10^5) and WMF (median:3.07 × 10^4), but lower in RD (median:3.60 × 10^2). The 25-75 % interquartile range of SARS-CoV2 copies in the NPS was significantly higher in patients who shed the virus in WMF (p = 0.0001) and RD (p = 0.01). Multivariate analyses showed that hospitalized patients shed significantly higher virus copies in the WMF (p = 0.01). Hospitalized patients with more severe disease (p = 0.03) and higher IL-6 values (p = 0.001) shed more SARS-CoV2 virus in the RD. CONCLUSIONS: WMF may be used reliably as a surrogate for diagnosis. High copy numbers in the NPS probably imply early disease onset, while in the WMF and RD may imply more severe disease and increased inflammation.


Subject(s)
Exhalation , Mouth/virology , Nasopharynx/virology , SARS-CoV-2/isolation & purification , Adult , COVID-19/diagnosis , COVID-19/virology , COVID-19 Nucleic Acid Testing , Cross-Sectional Studies , Female , Humans , Interleukin-6/blood , Male , Middle Aged , RNA, Viral/analysis , SARS-CoV-2/genetics , Sensitivity and Specificity , Severity of Illness Index , Viral Load , Virus Shedding
11.
Clin Exp Dent Res ; 7(5): 852-860, 2021 10.
Article in English | MEDLINE | ID: covidwho-1209677

ABSTRACT

OBJECTIVES: This pilot survey aims to study the oral manifestations associated with COVID-19 infection and report the prevalence of oral signs and symptoms in COVID-19 patients. MATERIALS AND METHODS: From May 15 to June 10, 2020, we used an online questionnaire containing the oral manifestations that are expected to be associated with the COVID-19 infection. Adults in our survey who have been diagnosed with COVID-19 positive were confirmed with reverse transcriptase PCR (RT-PCR), and isolated in various hospitals in Cairo, Egypt. RESULTS: This pilot study included 58 (53.4% males and 46.6% females) COVID-19 patients ages 18-46 years, and 13 (22.4%) were healthcare workers. Our results showed that 67.2% of the patients had at least one manifestation related to the oral cavity and salivary glands, and 32.8% (n = 19) did not have any symptoms associated with the oral cavity. The highest prevalence symptoms were dry mouth 39.7% (n = 23), gustatory dysfunction as 34.5% (n = 20) loss of salt sensation, 29.3% (n = 17) loss of sweet sensation, and 25.9% (n = 15) altered food taste, while the least prevalent symptoms were tongue redness 8.8% (n = 5), and gingival bleeding 7% (n = 4). The most frequently associated symptoms were loss of salt and sweetness, as reported by 27.6% of the participants. However, there was no significant association between the incidence of oral symptoms and demographic data (age, gender, or job) of the patients (p > 0.05). CONCLUSIONS: Based on limited data, COVID-19 significantly impacts the oral cavity and salivary glands, as salivary gland-related symptoms and taste disorders are highly prevalent in COVID-19 patients.


Subject(s)
COVID-19/complications , Mouth Diseases/epidemiology , Mouth/virology , SARS-CoV-2/isolation & purification , Adolescent , Adult , COVID-19/transmission , COVID-19/virology , Egypt/epidemiology , Female , Humans , Male , Middle Aged , Mouth Diseases/virology , Online Systems , Pilot Projects , Surveys and Questionnaires , Young Adult
12.
J Gen Virol ; 102(4)2021 04.
Article in English | MEDLINE | ID: covidwho-1207368

ABSTRACT

Infectious SARS-CoV-2 can be recovered from the oral cavities and saliva of COVID-19 patients with potential implications for disease transmission. Reducing viral load in patient saliva using antiviral mouthwashes may therefore have a role as a control measure in limiting virus spread, particularly in dental settings. Here, the efficacy of SARS-CoV-2 inactivation by seven commercially available mouthwashes with a range of active ingredients were evaluated in vitro. We demonstrate ≥4.1 to ≥5.5 log10 reduction in SARS-CoV-2 titre following a 1 min treatment with commercially available mouthwashes containing 0.01-0.02 % stabilised hypochlorous acid or 0.58 % povidone iodine, and non-specialist mouthwashes with both alcohol-based and alcohol-free formulations designed for home use. In contrast, products containing 1.5 % hydrogen peroxide or 0.2 % chlorhexidine gluconate were ineffective against SARS-CoV-2 in these tests. This study contributes to the growing body of evidence surrounding virucidal efficacy of mouthwashes/oral rinses against SARS-CoV-2, and has important applications in reducing risk associated with aerosol generating procedures in dentistry and potentially for infection control more widely.


Subject(s)
Antiviral Agents/pharmacology , Mouthwashes/pharmacology , SARS-CoV-2/drug effects , Virus Inactivation/drug effects , COVID-19/prevention & control , COVID-19/transmission , Cell Survival/drug effects , Humans , Mouth/virology , Viral Load/drug effects
13.
Nat Med ; 27(5): 892-903, 2021 05.
Article in English | MEDLINE | ID: covidwho-1152866

ABSTRACT

Despite signs of infection-including taste loss, dry mouth and mucosal lesions such as ulcerations, enanthema and macules-the involvement of the oral cavity in coronavirus disease 2019 (COVID-19) is poorly understood. To address this, we generated and analyzed two single-cell RNA sequencing datasets of the human minor salivary glands and gingiva (9 samples, 13,824 cells), identifying 50 cell clusters. Using integrated cell normalization and annotation, we classified 34 unique cell subpopulations between glands and gingiva. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral entry factors such as ACE2 and TMPRSS members were broadly enriched in epithelial cells of the glands and oral mucosae. Using orthogonal RNA and protein expression assessments, we confirmed SARS-CoV-2 infection in the glands and mucosae. Saliva from SARS-CoV-2-infected individuals harbored epithelial cells exhibiting ACE2 and TMPRSS expression and sustained SARS-CoV-2 infection. Acellular and cellular salivary fractions from asymptomatic individuals were found to transmit SARS-CoV-2 ex vivo. Matched nasopharyngeal and saliva samples displayed distinct viral shedding dynamics, and salivary viral burden correlated with COVID-19 symptoms, including taste loss. Upon recovery, this asymptomatic cohort exhibited sustained salivary IgG antibodies against SARS-CoV-2. Collectively, these data show that the oral cavity is an important site for SARS-CoV-2 infection and implicate saliva as a potential route of SARS-CoV-2 transmission.


Subject(s)
COVID-19/virology , Mouth/virology , SARS-CoV-2/isolation & purification , Saliva/virology , Angiotensin-Converting Enzyme 2/analysis , Asymptomatic Infections , COVID-19/etiology , Humans , Serine Endopeptidases/analysis , Taste Disorders/etiology , Taste Disorders/virology , Virus Replication
14.
Rev Med Virol ; 31(6): e2226, 2021 11.
Article in English | MEDLINE | ID: covidwho-1107716

ABSTRACT

The coronavirus disease 2019 (Covid-19) is a viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that clinically affects multiple organs of the human body. Cells in the oral cavity express viral entry receptor angiotensin-converting enzyme 2 that allows viral replication and may cause tissue inflammation and destruction. Recent studies have reported that Covid-19 patients present oral manifestations with multiple clinical aspects. In this review, we aim to summarise main signs and symptoms of Covid-19 in the oral cavity, its possible association with oral diseases, and the plausible underlying mechanisms of hyperinflammation reflecting crosstalk between Covid-19 and oral diseases. Ulcers, blisters, necrotising gingivitis, opportunistic coinfections, salivary gland alterations, white and erythematous plaques and gustatory dysfunction were the most reported clinical oral manifestations in patients with Covid-19. In general, the lesions appear concomitant with the loss of smell and taste. Multiple reports show evidences of necrotic/ulcerative gingiva, oral blisters and hypergrowth of opportunistic oral pathogens. SARS-CoV-2 exhibits tropism for endothelial cells and Covid-19-mediated endotheliitis can not only promote inflammation in oral tissues but can also facilitate virus spread. In addition, elevated levels of proinflammatory mediators in patients with Covid-19 and oral infectious disease can impair tissue homeostasis and cause delayed disease resolution. This suggests potential crosstalk of immune-mediated pathways underlying pathogenesis. Interestingly, few reports suggest recurrent herpetic lesions and higher bacterial growth in Covid-19 subjects, indicating SARS-CoV-2 and oral virus/bacteria interaction. Larger cohort studies comparing SARS-CoV-2 negative and positive subjects will reveal oral manifestation of the virus on oral health and its role in exacerbating oral infection.


Subject(s)
COVID-19/complications , Gingivitis, Necrotizing Ulcerative/complications , Herpesviridae Infections/complications , Oral Ulcer/complications , Periodontal Diseases/complications , Sialadenitis/complications , Stomatitis, Aphthous/complications , Xerostomia/complications , Angiotensin-Converting Enzyme 2/genetics , Angiotensin-Converting Enzyme 2/immunology , Anosmia/complications , Anosmia/immunology , Anosmia/pathology , Anosmia/virology , COVID-19/immunology , COVID-19/pathology , COVID-19/virology , Dysgeusia/complications , Dysgeusia/immunology , Dysgeusia/pathology , Dysgeusia/virology , Gene Expression , Gingivitis, Necrotizing Ulcerative/immunology , Gingivitis, Necrotizing Ulcerative/pathology , Gingivitis, Necrotizing Ulcerative/virology , Herpesviridae Infections/immunology , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Humans , Mouth/immunology , Mouth/pathology , Mouth/virology , Oral Ulcer/immunology , Oral Ulcer/pathology , Oral Ulcer/virology , Periodontal Diseases/immunology , Periodontal Diseases/pathology , Periodontal Diseases/virology , SARS-CoV-2/immunology , SARS-CoV-2/pathogenicity , Serine Endopeptidases/genetics , Serine Endopeptidases/immunology , Sialadenitis/immunology , Sialadenitis/pathology , Sialadenitis/virology , Stomatitis, Aphthous/immunology , Stomatitis, Aphthous/pathology , Stomatitis, Aphthous/virology , Xerostomia/immunology , Xerostomia/pathology , Xerostomia/virology
15.
PLoS One ; 15(10): e0241542, 2020.
Article in English | MEDLINE | ID: covidwho-1067389

ABSTRACT

Oral swabs are emerging as a non-invasive sample type for diagnosing infectious diseases including Ebola, tuberculosis (TB), and COVID-19. To assure proper sample collection, sample adequacy controls (SACs) are needed that detect substances indicative of samples collected within the oral cavity. This study evaluated two candidate SACs for this purpose. One detected representative oral microbiota (Streptococcus species DNA) and the other, human cells (human mitochondrial DNA, mtDNA). Quantitative PCR (qPCR) assays for the two target cell types were applied to buccal swabs (representing samples collected within the oral cavity) and hand swabs (representing improperly collected samples) obtained from 51 healthy U.S. volunteers. Quantification cycle (Cq) cutoffs that maximized Youden's index were established for each assay. The streptococcal target at a Cq cutoff of ≤34.9 had 99.0% sensitivity and specificity for oral swab samples, whereas human mtDNA perfectly distinguished between hand and mouth swabs with a Cq cutoff of 31.3. The human mtDNA test was then applied to buccal, tongue, and gum swabs that had previously been collected from TB patients and controls in South Africa, along with "air swabs" collected as negative controls (total N = 292 swabs from 71 subjects). Of these swabs, 287/292 (98%) exhibited the expected Cq values. In a paired analysis the three oral sites yielded indistinguishable amounts of human mtDNA, however PurFlockTM swabs collected slightly more human mtDNA than did OmniSwabsTM (p = 0.012). The results indicate that quantification of human mtDNA cannot distinguish swabs collected from different sites within the mouth. However, it can reliably distinguish oral swabs from swabs that were not used orally, which makes it a useful SAC for oral swab-based diagnosis.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , SARS-CoV-2/isolation & purification , Specimen Handling/methods , Adult , COVID-19/epidemiology , COVID-19/transmission , COVID-19/virology , DNA, Mitochondrial/analysis , DNA, Mitochondrial/genetics , DNA, Viral/analysis , DNA, Viral/genetics , Diagnostic Tests, Routine/methods , Female , Humans , Male , Mouth/virology , Real-Time Polymerase Chain Reaction , Reference Standards , Sensitivity and Specificity , South Africa/epidemiology , Washington/epidemiology
16.
Bol Med Hosp Infant Mex ; 78(2): 91-94, 2021 01 26.
Article in English | MEDLINE | ID: covidwho-1049284

ABSTRACT

Coronavirus disease 2019 (COVID-19) is a new disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. China reported the first case of COVID-19 in December 2019, and a few months later, the World Health Organization declared it as a pandemic. Oral ulcers in adult patients have been associated with COVID-19. However, no cases have yet been documented in children. The angiotensin-converting enzyme-2 (ACE2) receptor has been identified in tissues of the oral cavity. Studies have identified the tongue as the site with the highest expression of ACE2, and the oral epithelium, gingival epithelium, and salivary glands as sites of lesser extent expression. ACE2 expression is lower in children and varies with age. SARS-CoV-2 in saliva has been identified in various studies, which suggests that this could be a useful sample for diagnosis. However, its presence in saliva would indicate the high risk of contagion of this fluid.


Subject(s)
COVID-19/complications , Oral Health , Oral Ulcer/virology , SARS-CoV-2/isolation & purification , Adult , Age Factors , Angiotensin-Converting Enzyme 2/metabolism , COVID-19/diagnosis , COVID-19/virology , COVID-19 Testing , Child , Humans , Mouth/virology , Saliva/virology
18.
Virology ; 554: 97-105, 2021 02.
Article in English | MEDLINE | ID: covidwho-1003123

ABSTRACT

We examined the pathogenicity of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in cynomolgus macaques for 28 days to establish an animal model of COVID-19 for the development of vaccines and antiviral drugs. Cynomolgus macaques infected with SARS-CoV-2 showed body temperature rises and X-ray radiographic pneumonia without life-threatening clinical signs of disease. A neutralizing antibody against SARS-CoV-2 and T-lymphocytes producing interferon (IFN)-γ specifically for SARS-CoV-2 N-protein were detected on day 14 in one of three macaques with viral pneumonia. In the other two macaques, in which a neutralizing antibody was not detected, T-lymphocytes producing IFN-γ specifically for SARS-CoV-2 N protein increased on day 7 to day 14, suggesting that not only a neutralizing antibody but also cellular immunity has a role in the elimination of SARS-CoV-2. Thus, because of similar symptoms to approximately 80% of patients, cynomolgus macaques are appropriate to extrapolate the efficacy of vaccines and antiviral drugs for humans.


Subject(s)
Antibodies, Neutralizing/immunology , COVID-19/immunology , Disease Models, Animal , SARS-CoV-2/immunology , T-Lymphocytes/immunology , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Antibodies, Viral/immunology , COVID-19/pathology , COVID-19/virology , Cytokines/blood , Female , Interferon-gamma/immunology , Macaca fascicularis , Male , Mouth/virology , Nasal Cavity/virology , Pneumonia, Viral/immunology , Pneumonia, Viral/pathology , Pneumonia, Viral/virology , SARS-CoV-2/pathogenicity , SARS-CoV-2/physiology , Viral Load
19.
PLoS One ; 15(12): e0243708, 2020.
Article in English | MEDLINE | ID: covidwho-999825

ABSTRACT

To slow the progression of COVID-19, the Centers for Disease Control (CDC) and the World Health Organization (WHO) have recommended wearing face coverings. However, very little is known about how occluding parts of the face might impact the emotion inferences that children make during social interactions. The current study recruited a racially diverse sample of school-aged (7- to 13-years) children from publicly funded after-school programs. Children made inferences from facial configurations that were not covered, wearing sunglasses to occlude the eyes, or wearing surgical masks to occlude the mouth. Children were still able to make accurate inferences about emotions, even when parts of the faces were covered. These data suggest that while there may be some challenges for children incurred by others wearing masks, in combination with other contextual cues, masks are unlikely to dramatically impair children's social interactions in their everyday lives.


Subject(s)
COVID-19/epidemiology , COVID-19/prevention & control , Emotions , Masks , Adolescent , COVID-19/psychology , COVID-19/virology , Child , Face , Facial Expression , Female , Humans , Male , Mouth/virology , SARS-CoV-2/pathogenicity , Social Interaction
20.
Emerg Infect Dis ; 27(1): 104-112, 2021 01.
Article in English | MEDLINE | ID: covidwho-993258

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the agent that causes coronavirus disease, has been shown to infect several species. The role of domestic livestock and associated risks for humans in close contact with food production animals remains unknown for many species. Determining the susceptibility of pigs to SARS-CoV-2 is critical to a One Health approach to manage potential risk for zoonotic transmission. We found that pigs are susceptible to SARS-CoV-2 after oronasal inoculation. Among 16 animals, we detected viral RNA in group oral fluids and in nasal wash from 2 pigs, but live virus was isolated from only 1 pig. Antibodies also were detected in only 2 animals at 11 and 13 days postinoculation but were detected in oral fluid samples at 6 days postinoculation, indicating antibody secretion. These data highlight the need for additional livestock assessment to determine the potential role of domestic animals in the SARS-CoV-2 pandemic.


Subject(s)
Antibodies, Viral/blood , Coronavirus Infections/veterinary , Coronavirus Infections/virology , RNA, Viral/blood , SARS-CoV-2/immunology , Animals , Antibodies, Neutralizing/blood , Disease Susceptibility/veterinary , Female , Lymph Nodes/virology , Male , Mouth/virology , Nasal Cavity/virology , Rectum/virology , SARS-CoV-2/isolation & purification , SARS-CoV-2/physiology , Swine , Virus Shedding
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