scholarly journals Molecular Screening and Analysis Reveal Novel Oral Site-Specific Locations for the Cariogenic Pathogen Scardovia wiggsiae

2021 ◽  
Vol 9 (6) ◽  
pp. 73
Author(s):  
Steven McDaniel ◽  
Jaydene McDaniel ◽  
Katherine M. Howard ◽  
Karl Kingsley
Keyword(s):  
Gingival Crevicular Fluid ◽  
Quantitative Polymerase Chain Reaction ◽  
Orthodontic Brackets ◽  
Oral Disease ◽  
Significant Advance ◽  
Clinical Samples ◽  
Site Specific ◽  
Clinical Patient ◽  
Specific Analysis ◽  
Oral Site

Introduction: Scardovia wiggsiae (SW) is a newly identified cariogenic pathogen associated with severe early childhood caries and oral disease. New studies have confirmed the presence of this organism among clinical samples from both pediatric and adult patients. However, the recent discovery of this organism has left researchers with only limited information available regarding the prevalence of this organism—and virtually no information regarding oral site-specific locations. Based upon this lack of information, the overall objective of this study was to perform an oral site-specific analysis of SW prevalence from clinical samples. Methods: Using an approved human subjects protocol, samples (n = 60) from an existing saliva and site-specific biorepository were identified and screened for SW presence using quantitative polymerase chain reaction (qPCR). These data were summarized and subsequently analyzed for correlations with demographic (age, sex, race or ethnicity) or clinical (body mass index or BMI, primary/mixed/permanent dentition, orthodontic brackets) variables. Results: These data revealed that average DNA concentrations from all sample sites (saliva, dorsum of tongue, gingival crevicular fluid (GCF), biofilm of upper buccal molar, and biofilm of lower lingual incisor) ranged between 13.74 and 14.69 μg/μL, with an overall average of 14.30 μg/μL ± 1.12 (standard error or SE). qPCR screening revealed a total of n = 34/60 or 56.7% of patient samples harboring SW. A total of n = 71/170 specific oral sites harbored this organism, with the majority of the SW-positive participant samples harboring SW at more than one oral site, n = 22/34 or 64.7%, including non-traditional sites such as GCF and the dorsum of the tongue. Weak correlations were found between specific SW outcomes in GCF and type of dentition (permanent; R = 0.2444), as well as SW outcomes in saliva with age (R = 0.228) and presence of orthodontic brackets (R = 0.2118). Conclusions: This study may be among the first to provide oral site-specific analysis to reveal the prevalence and location of Scardovia among clinical patient samples. Moreover, these data also provide some of the first evidence to suggest this organism may be present not only in traditional supragingival tooth-associated biofilm sites, but also in non-traditional oral sites including the dorsum of the tongue and the gingival crevice. Based upon these results, these data may represent a significant advance in our understanding of the potential sites and locations that harbor this organism and may help contribute to our understanding of the prevalence, distribution and potential for the development of oral disease among clinic patients.

scholarly journals Oral Site Specific Sampling Reveals Differential Location for Scardovia wiggsiae

2020 ◽  
pp. 47-55
Author(s):  
Graydon Carr ◽  
Arvin Alexander ◽  
Linh Nguyen ◽  
Karl Kingsley
Keyword(s):  
Pcr Primers ◽  
Primary Objective ◽  
Oral Disease ◽  
Clinical Samples ◽  
Site Specific ◽  
New Information ◽  
Oral Site ◽  
Tooth Surfaces ◽  
High Risk Populations ◽  
Unstimulated Saliva

Introduction: The newly discovered cariogenic pathogen Scardovia wiggsiae has been the subject of intense scientific interest due to the role it may play in the development or progression of caries and oral disease. The primary objective of this study was to perform DNA microbial screening from five specific oral sites, including the gingival crevice between the upper central incisors, biofilm of the upper first molar and lingual incisor, as well as the dorsum of the tongue – for comparison with unstimulated saliva. These data may provide significant insights into site-specific oral locations that harbor S. wiggsiae. Methods: More than one hundred previously collected clinical samples (n=105) were identified for inclusion in this study. DNA isolates were screened using a NanoDrop spectrophotometer to determine overall DNA quantity and quality. Samples with sufficient quality and quantity were screened for the presence of S. wiggsiae using validated PCR primers. Results: More than one hundred patient samples (n=105) were identified, which were comprised of mostly female (57%) versus male (43%) and minority (71%) versus White (29%). The average DNA concentrations ranged between 13.74 and 14.69 mg/mL, with A260:A280 ratios ranging between 1.62 – 1.70. Results of molecular screening using S. wiggsiae specific primers demonstrated only a small percentage of pooled samples (7.6%) harbored this DNA, which was highly concentrated among the samples from tooth surfaces (Upper First Molar, Lingual Incisor) and saliva compared with the gingival crevice and dorsum of the tongue. Discussion: These data provide novel information regarding specific oral locations, including tooth surfaces that harbor S. wiggsiae. In addition, these sites also provide new information regarding oral sites that do not appear to harbor this organism, including the gingival crevice and dorsum of the tongue. This information may be particularly useful to oral health researchers as they strive to limit and reduce the cariogenic microbiome among high-risk populations.

scholarly journals Microbial Screening Reveals Oral Site-Specific Locations of the Periodontal Pathogen Selenomonas noxia

2021 ◽  
Vol 43 (1) ◽  
pp. 353-364
Author(s):  
Jaydene McDaniel ◽  
Steven McDaniel ◽  
Beanca Jhanine Samiano ◽  
Matthew Marrujo ◽  
Karl Kingsley ◽  
...  
Keyword(s):  
Low Socioeconomic Status ◽  
Gingival Crevicular Fluid ◽  
Clinical Information ◽  
Periodontal Pathogen ◽  
Site Specific ◽  
Low Socioeconomic ◽  
Oral Site ◽  
Crevicular Fluid ◽  
Selenomonas Noxia ◽  
Positive Results

Introduction: Selenomonas noxia (SN) is an important periodontal pathogen, associated with gingivitis and periodontitis. Many studies have found associations between SN and indicators of poor health outcomes, such as smoking, low socioeconomic status and obesity. However, less is known about the prevalence of this organism and more specifically about other oral site-specific locations that may harbor this organism. Methods: Using an existing patient repository (n = 47) of DNA isolated from saliva and other oral sites (n = 235), including the dorsum of the tongue, lower lingual incisor, upper buccal molar and gingival crevicular fluid (GCF), molecular screening for SN was performed. Screening results were analyzed for associations between demographic variables (age, sex, race/ethnicity) and clinical information (body mass index or BMI, presence of orthodontic brackets, primary/mixed/permanent dentition). Results: qPCR screening revealed a total of n = 62/235 sites or 26.3% harboring SN with saliva and GCF (either alone or in combination with one or more sites) most often observed (Saliva, n = 23/27 or 85.18%, GCF, n = 14/27 or 51%). Analysis of site-specific data revealed most positive results were found among saliva and GCF alone or in combination, with fewer positive results observed among the tongue (33.3%), lower lingual incisor (29.6%), and upper buccal molar (25.9%). No significant associations were found between demographic or clinical variables and presence of SN at any site. Conclusions: These results may be among the first to describe site-specific locations of S. noxia among various additional oral biofilm sites. These data may represent a significant advancement in our understanding of the sites and locations that harbor this organism, which may be important for our understanding of the prevalence and distribution of these organisms among patients of different ages undergoing different types of oral treatments, such as orthodontic treatment or therapy.

scholarly journals Development and validation of a multiplex qPCR assay for detection and relative quantification of HPV16 and HPV18 E6 and E7 oncogenes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexia Bordigoni ◽  
Anne Motte ◽  
Hervé Tissot-Dupont ◽  
Philippe Colson ◽  
Christelle Desnues
Keyword(s):  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Papillomaviruses ◽  
Molecular Techniques ◽  
Clinical Samples ◽  
Gene Encoding ◽  
Hpv Dna ◽  
E6 And E7 ◽  
High Risk Hpv ◽  
Development And Validation

AbstractHuman papillomaviruses (HPV) play a key role in promoting human anogenital cancers. Current high-risk HPV screening or diagnosis tests involve cytological or molecular techniques mostly based on qualitative HPV DNA detection. Here, we describe the development of a rapid quantitative polymerase chain reaction (qPCR) detection test of HPV16 and HPV18 oncogenes (E6 and E7) normalized on human gene encoding GAPDH. Optimized qPCR parameters were defined, and analytical specificities were validated. The limit of detection was 101 for all genes tested. Assay performances were evaluated on clinical samples (n = 96). Concordance between the Xpert HPV assay and the triplex assay developed here was 93.44% for HPV16 and 73.58% for HPV18. HPV co-infections were detected in 15 samples. The systems developed in the present study can be used in complement to traditional HPV tests for specifically validating the presence of HPV16 and/or HPV18. It can also be used for the follow-up of patients with confirmed infection and at risk of developing lesions, through the quantification of E6 and E7 oncogene expression (mRNA) normalized on the GAPDH expression levels.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

A new ultrasensitive chemiluminescent assay for the site-specific quantification of alkaline phosphatase in gingival crevicular fluid

1993 ◽  
Vol 28 (4) ◽  
pp. 266-273 ◽  
Author(s):  
I. L. C. Chapple ◽  
J. B. Matthews ◽  
G. H. G. Thorpe ◽  
H. D. Glenwright ◽  
J. M. Smith ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Gingival Crevicular Fluid ◽  
Site Specific ◽  
Crevicular Fluid

scholarly journals Quantitative Chemoproteomics for Site-Specific Analysis of Protein Alkylation by 4-Hydroxy-2-Nonenal in Cells

2015 ◽  
Vol 87 (5) ◽  
pp. 2535-2541 ◽  
Author(s):  
Jing Yang ◽  
Keri A. Tallman ◽  
Ned A. Porter ◽  
Daniel C. Liebler
Keyword(s):  
Site Specific ◽  
Specific Analysis ◽  
In Cells

scholarly journals Global site-specific analysis of glycoprotein N-glycan processing

2018 ◽  
Vol 13 (6) ◽  
pp. 1196-1212 ◽  
Author(s):  
Liwei Cao ◽  
Jolene K Diedrich ◽  
Yuanhui Ma ◽  
Nianshuang Wang ◽  
Matthias Pauthner ◽  
...  
Keyword(s):  
Site Specific ◽  
Specific Analysis
Sign in / Sign up

Export Citation Format

Share Document