Comparison of the Performance of the PanBio COVID-19 Antigen Test in SARS-CoV-2 B.1.1.7 (Alpha) Variants versus non-B.1.1.7 Variants.

This study evaluates the performance of the PanBio COVID-19 antigen (Ag) test as part of a hospital infection control policy. Hospital staff was encouraged to get tested for COVID-19 when presenting with SARS-CoV-2-related symptoms. In a period of approximately 5 months, a steady decline in the performance of the Ag test was noted, epidemiologically coinciding with the rise of the SARS-CoV-2 B.1.1.7 (alpha) variant of concern (VOC) in the Netherlands. This led to the hypothesis that the diagnostic performance of the PanBio COVID-19 Ag test was influenced by the infecting viral variant. The results show a significantly lower sensitivity of the PanBio COVID-19 Ag test in persons infected with the B.1.1.7 (alpha) variant of SARS-CoV-2 in comparison with that in persons infected with non-B.1.1.7 variants, also after adjustment for viral load. IMPORTANCE Antigen tests for COVID-19 are widely used for rapid identification of COVID-19 cases, for example, for access to schools, festivals, and travel. There are several FDA- and CE-cleared tests on the market. Their performance has been evaluated mainly on the basis of infections by the classical variant of the causing virus, SARS-CoV-2. This paper provides evidence that the performance of one of the most widely used antigen tests detects significantly fewer cases of COVID-19 by the alpha variant than by the classical variants of SARS-CoV-2. This means that the role of antigen tests needs to be reevaluated in regions where other variants of SARS-CoV-2 predominate.

Feasibility of a quantitative polymerase chain reaction assay for diagnosing pneumococcal pneumonia using oropharyngeal swabs.

Streptococcus pneumoniae is the most important pathogen causing community-acquired pneumonia (CAP). The current diagnostic microbial standard detects S. pneumoniae in less than 30% of CAP cases. A quantitative polymerase chain reaction (PCR) targeting autolysin (lytA) is able to increase the rate of detection. The aim of this study is validation of this quantitative PCR in vitro using different available strains and in vivo using clinical samples (oropharyngeal swabs). The PCR autolysin (lytA) was validated by testing the intra- and inter-run variability. Also, the in vitro specificity and sensitivity, including the lower limit of detection was determined. In addition, a pilot-study was performed using samples from patients (n = 28) with pneumococcal pneumonia and patients (n = 28) with a pneumonia without detection of S. pneumoniae with the current diagnostic microbial standard, but with detection of either a viral and or another bacterial pathogen to validate this test further. The intra- and inter-run variability were relatively low (SD's ranging from 0.08 to 0.96 cycle thresholds). The lower limit of detection turned out to be 1-10 DNA copies/reaction. In-vitro sensitivity and specificity of the tested specimens (8 strains carrying lytA and 6 strains negative for lytA) were both 100%. In patients with pneumococcal and non-pneumococcal pneumonia a cut-off value of 6.000 copies/mL would lead to a sensitivity of 57.1% and a specificity of 85.7%. We were able to develop a quantitative PCR targeting lytA with good in-vitro test characteristics.

Case-control comparison of bacterial and protozoan microorganisms associated with gastroenteritis: application of molecular detection.

The introduction of molecular detection of infectious organisms has led to increased numbers of positive findings, as observed for pathogens causing gastroenteritis (GE). However, because little is known about the prevalence of these pathogens in the healthy asymptomatic population, the clinical value of these additional findings is unclear. A case-control study was carried out in a population of patients served by general practitioners in the Netherlands. A total of 2710 fecal samples from case and matched control subjects were subjected to multiplex real-time PCR for the 11 most common bacterial and four protozoal causes of GE. Of 1515 case samples, 818 (54%) were positive for one or more target organisms. A total of 49% of the controls were positive. Higher positivity rates in cases compared to controls were observed for Campylobacter spp., Salmonella spp., Clostridium difficile, enteroinvasive Escherichia coli/Shigella spp., enterotoxigenic E. coli, enteroaggregative E. coli, atypical enteropathogenic E. coli (EPEC), Cryptosporidium parvum/hominis, and Giardia lamblia. However, Dientamoeba fragilis and Shiga-like toxigenic E. coli were detected significantly less frequent in cases than in controls, while no difference in prevalence was found for typical EPEC and enterohemorrhagic E. coli. The association between the presence of microorganisms and GE was the weakest in children aged 0 to 5 years. Higher relative loads in cases further support causality. This was seen for Campylobacter spp., Salmonella spp., enterotoxigenic E. coli, and C. parvum/hominis, and for certain age categories of those infected with C. difficile, enteroaggregative E. coli, and atypical EPEC. For D. fragilis and Shiga-like toxigenic E. coli/enterohemorrhagic E. coli, pathogen loads were lower in cases. Application of molecular diagnostics in GE is rapid, sensitive and specific, but results should be interpreted with care, using clinical and additional background information.

Comparison of periodontal pathogens between cats and their owners.

The periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia are strongly associated with periodontal disease and are highly prevalent in humans with periodontitis. Porphyromonas and Tannerella spp. have also been isolated from the oral cavity of cats. The oral microflora in animals was compared with those in humans in earlier studies, but no studies are available on the comparison of the oral microflora from pets and their respective owners. The aim of this study was to determine the presence of these bacteria in the oral microflora of cats and their owners, since animal to human transmission, or vice versa, of oral pathogens could have public health implications. This study investigated the prevalence of Porphyromonas gulae, P. gingivalis, and T. forsythia in the oral microflora of cats and their owners, using culture and polymerase chain reaction (PCR). All Porphyromonas isolates from cats (n=64) were catalase positive, whereas the Porphyromonas isolates from owners (n=7) were catalase negative, suggesting that the isolates from cats were P. gulae whereas those from the owners were P. gingivalis. T. forsythia was recovered from both cats (n=63) and owners (n=31); the proportion of T. forsythia relative to the total CFU was higher in cats with periodontitis than in cats without periodontal disease. Genotyping of T. forsythia isolates (n=54) in six cat/owner couples showed that in one cat/owner couple the T. forsythia isolates (n=6) were identical. These T. forsythia isolates were all catalase positive, which led us to hypothesize that transmission from cats to owners had occurred and that cats may be a reservoir of T. forsythia.

[Stress and periodontal health]. / Stress en parodontale gezondheid.

Periodontitis and periimplant infections are complex manifestations associated with several disease-modifying factors, such as causative pathogens and smoking. Although research into these factors has led to important progressions in the treatment of these infections in recent decades, the contribution of mental stress in the absence of pathogens or smoking is still unclear. Qualitative and quantitative assessment of mental stress might be an important instrument in periodontal and periimplant therapy.

Porphyromonas gingivalis, Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction.

BACKGROUND AND AIMS: Bacteria play an essential role in the pathogenesis of destructive periodontal disease. It has been suggested that not all bacteria associated with periodontitis may be normal inhabitants of a periodontally healthy dentition. In particular, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans have been isolated infrequently from subjects without periodontitis. The aim of the present study was to compare prevalence and proportions of a number of periodontal bacteria in periodontitis patients and control subjects. MATERIAL AND METHODS: In all, 116 consecutive subjects diagnosed with moderate to severe periodontitis (mean age 42.4) and 94 subjects without radiographic evidence of alveolar bone loss (mean age 40.4) were recruited for the study. The gingival condition in the control group varied between gingival health and various degrees of gingivitis. In patients, the deepest pocket in each quadrant was selected for microbiological sampling. In control subjects all mesial and distal sites of all first molars were selected for sampling. All paper points from a patient were pooled and processed for anaerobic cultivation within 6 h after sampling. Clinical variables of sampled sites included bleeding index, probing pocket depth and clinical attachment level. RESULTS: A. actinomycetemcomitans, P. gingivalis, Prevotella intermedia, Bacteroides forsythus, Fusobacterium nucleatum and Peptostreptococcus micros were significantly more often prevalent in patients than in controls. The highest odds ratios were found for P. gingivalis and B. forsythus (12.3 and 10.4 resp.). Other odds ratios varied from 3.1 to 7.7 for A. actinomycetemcomitans and P. micros, respectively. Absolute numbers of target bacteria were all higher in patients, but only the mean percentage of B. forsythus was significantly higher in patients in comparison to controls (P < 0.001). CONCLUSIONS: A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus, F. nucleatum and P. micros are all significant markers for destructive periodontal disease in adult subjects. Based on calculated odds ratios, B. forsythus and P. gingivalis are the strongest bacterial markers for this disease and are infrequently cultured from subjects without periodontal bone loss.

Absence of a specific subgingival microflora in adults with Down's syndrome.

BACKGROUND: Periodontal disease in Down's syndrome (DS) is generally characterized by a high degree of bone loss. Bone loss of 5 mm or more is observed in 70% of these subjects. Among DS subjects, considerable differences in disease progression occur. So far, no studies have been conducted in which specific properties of the subgingival microflora have been related to the condition observed. AIMS: To investigate (1) the subgingival microflora in DS subjects and other mentally retarded (control) individuals which were matched to the utmost and (2) to investigate the subgingival microflora of a "low-risk" and a " high-risk" group formed in DS subjects. MATERIAL AND METHODS: 17 DS subjects and 17 control subjects were matched with respect to age, plaque level and bleeding on probing. In addition, the DS group was divided in a "low-risk" group (0-2 teeth lost due to periodontal disease n=6) and a "high-risk"group (6-13 teeth lost due to periodontal disease n=11). Prevalence and proportions of the putative periodontal pathogens Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Peptostreptococcus micros, Fusobacterium nucleatum and Campylobacter rectus in the subgingival plaque were determined using anaerobic culture techniques. No differences in the prevalence of distinct suspected periodontopathic bacteria and bacterial subgingival composition between the DS group and the control group could be established. Also no differences in the prevalence of the seven investigated microbial species between the "low-risk" and the "high-risk" group were observed. CONCLUSIONS: Because of the lack of differences in microflora between the DS group and the control group, a specific effect of the microbiological composition in the periodontal status of subjects with DS can be excluded in this population. Host factors constitute the more likely explanation of the differences observed in DS.

Mutans streptococci in subgingival plaque of treated and untreated patients with periodontitis.

BACKGROUND: The etiology of root caries is thought to be identical to coronal caries, though root caries seem to be more complicated because of the higher susceptibility of exposed roots (dentin) by periodontal therapy to demineralization than intact enamel. This implies that mutans streptococci are the most likely pathogens in the development of root caries. Although it is known that both the numbers of mutans streptococci and the frequency of isolation in root caries lesions are negatively correlated with the distance from the gingival margin, the subgingival sulcus has not been considered a possible habitat for mutans streptococci. However, subgingival occurrence of mutans streptococci in both untreated and treated periodontal patients has not been documented well in the literature. OBJECTIVE: To investigate the presence and levels of mutans streptococci in the subgingival plaque of patients (n=154) in different stages of periodontal therapy. The subgingival sulcus may be a possible habitat for mutans streptococci. This localisation of mutans streptococci may be of importance in the development of root caries after periodontal surgery. MATERIALS AND METHODS: In this cross-sectional study, subgingival plaque samples from 154 consecutive adult periodontitis patients were tested for presence and levels of mutans streptococci and putative periodontal pathogens by anaerobic cultures. These patients were divided into 4 groups based on their stage of periodontal treatment: (1) untreated patients; (2) patients after initial periodontal therapy only; (3) patients in the maintenance phase who not underwent periodontal surgery; (4) patients after periodontal surgery. RESULTS: The prevalence of mutans streptococci in the 4 study groups varied from 82% in untreated patients to 94% in maintenance patients. The mean proportion of mutans streptococci was 6.65% in maintenance patients versus 1.86% in untreated patients (p=0.005) and 2.51% in patients after scaling and root planing (p=0.041). CONCLUSIONS: The subgingival area is a microbial habitat for mutans streptococci that may be of importance in the development of root caries in periodontitis patients.

Smoking affects the subgingival microflora in periodontitis.

BACKGROUND: Tobacco smoking has been identified as one major risk factor for destructive periodontal disease. Scaling and root planing have been shown to be less effective in smokers with periodontitis. The aim of the present study was to compare the subgingival microbial flora of treated and untreated smokers and non-smokers. METHODS: Four independent adult patient groups with periodontitis were included in this investigation: 88 untreated smokers (U-S); 90 untreated non-smokers (U-NS); 119 treated non-smokers (T-NS); and 171 treated smokers (T-S). Clinical variables included cumulative plaque index (CPI), probing depth (PD), clinical attachment level (CAL), cumulative bleeding index (CBI), and cumulative suppuration index (CSI). Paper point samples from the deepest bleeding pocket in each quadrant of the dentition were analyzed for the presence and levels of 6 periodontal bacterial pathogens using anaerobic culture techniques. RESULTS: U-S showed a higher mean cumulative plaque index than U-NS (3.5 versus 2.7). Mean PD and mean CAL were higher in the T-S in comparison to the T-NS group (7.0 versus 6.6 mm and 5.6 versus 4.7 mm, respectively). Microbiological characteristics of U-S were a higher prevalence of Prevotella intermedia/nigrescens and higher mean levels of Peptostreptococcus micros (Pm) and Fusobacterium nucleatum (Fn). T-S patients were characterized by higher prevalence of Bacteroides forsythus (Bf), Pm, and Campylobacter rectus (Cr) and higher mean levels of Pm and Fn. The mean percentage of B. forsythus tended to be higher in the T-S group than in the T-NS group (6.9% versus 5.6%). The relative risk to be infected with Bf, Pm, and Cr was statistically higher in smokers (odds ratios: 1.9, 1.9, and 1.6, respectively). The chance to find > or =10% of Bf, Pm, and/or Fn was 3.3 higher in smokers when A. actinomycetemcomitans and P gingivalis were absent. Detection of > or =20% Pm/Fn in treated patients was strongly associated with smoking (odds ratio 13.8, P= 0.002). CONCLUSIONS: Smoking is a determining factor for the composition of the subgingival microflora in adult patients with periodontitis and may select for a specific cluster of periodontal pathogens, notably Bf, Pm, Fn, and Cr. On the basis of these observations, smoking, among other criteria, may be one parameter to use in deciding to treat refractory periodontitis in smokers with a systemic antibiotic therapy directed against smoking-associated periodontal bacteria.

Evaluation of the use of xanthan as vehicle for cationic antifungal peptides.

Oral candidiasis frequently occurs in individuals with dry mouth syndrome (xerostomia), in immunocompromised patients and in denture wearers. The aim of this study was to develop a formulation which will prolong the retention time of antimicrobial agents at the site of application. The activity against Candida albicans of a synthetic cationic peptide dhvar 1, based on the human fungicidal salivary peptide histatin 5, was tested either in a mixture with the bioadhesive polymer xanthan, or after covalent coupling to this polymer. The presence of xanthan resulted in an increase of the LC50 value of the peptide from 2.6 (S.D.=0.6) to 5.8 (S.D.=4.0). Covalent coupling caused an additional increase of the LC50 value to 18.4 (S. D.=6.7). Coupling caused a reduction of the viscosity and elasticity of the xanthan solution related to the applied concentration of the coupling agent. Incubation of the peptide with clarified human whole saliva resulted in proteolytic degradation of the peptide. In the presence of xanthan the degradation occurred more slowly. It was concluded that xanthan is an appropriate vehicle for antimicrobial peptides in a retention increasing formulation.

Influence of polymers for use in saliva substitutes on de- and remineralization of enamel in vitro.

A number of polymers which have previously been tested for their applicability as thickening agents in saliva substitutes were studied in vitro for their caries-protective properties. These were: polyacrylic acid, carboxymethylcellulose, xanthan gum, guar gum, hydroxyethylcellulose and porcine gastric mucin. The polymers were tested for their effects on: (1) growth of hydroxyapatite crystals in a supersaturated calcium phosphate solution, (2) dissolution of hydroxyapatite crystals in 50 mM acetic acid, pH 5.2 and (3) demineralization and remineralization of bovine enamel in a pH-cycling model. Growth of hydroxyapatite crystals was strongly inhibited by polyacrylic acid and carboxymethylcellulose at very low concentrations (0.005% w/v). Other polymers displayed lower inhibition of hydroxyapatite crystal growth. Hydroxyapatite dissolution was inhibited by all polymers except by hydroxymethylcellulose and xanthan gum. This occurred both in the presence of the polymers as well as after a 30-min preincubation. In the pH-cycling experiment, bovine enamel specimens with preformed lesions were alternately exposed to a demineralization buffer and a remineralization buffer containing the polymers hydroxyethylcellulose, carboxymethylcellulose, xanthan gum, polyacrylic acid, or porcine gastric mucin. A remineralization buffer containing 1 ppm NaF was used as a positive control. Under the experimental conditions, the control experiment without additives resulted in a net mineral loss (30.6 mumol Ca/cm2 after 14 days of pH cycling). In the presence of 1 ppm NaF, a small mineral gain was observed (8.6 mumol/cm2). All polymers largely inhibited further demineralization (1.2-12.3 mumol/cm2) except polyacrylic acid which, inhibited of its high calcium-binding capacity, caused demineralization, especially in the remineralization buffer (17.1 mumol/cm2). In conclusion, polymers tested in this study, except the polyacrylic acid, reduced the demineralization of enamel in vitro. The precise mechanism of the protective effect is not clear but it is speculated that formation of an absorbed polymer layer on the hydroxyapatite or enamel surface may provide protection against acidic attacks.

Analysis of the concentration and output of whole salivary constituents in patients with Sjögren's syndrome.

In Sjögren's syndrome, salivary glands are affected, resulting in a diminished salivary flow. In the present study, the protein composition, sialic acid content and the amounts of calcium and phosphate of stimulated whole saliva from 43 patients with Sjögren's syndrome, were compared with those of control saliva samples from 17 healthy subjects. The absolute concentrations of albumin, cystatin C, cystatin S, total IgA and total protein, but not amylase, were increased significantly in both primary and secondary Sjögren's syndrome. The output/min of total protein, albumin, amylase, and IgA was, however, decreased in Sjögren patients. These results suggest that the diminished output of salivary defence factors, rather than their absolute concentrations, may be related to the oral health problems seen in Sjögren's syndrome patients.

Treatment of xerostomia with polymer-based saliva substitutes in patients with Sjögren's syndrome.

OBJECTIVE: To determine the efficacy of 3 types of polymer-based saliva substitutes in reducing oral dryness in patients with Sjögren's syndrome (SS). METHODS: Subjective efficacy of 3 different saliva substitutes (determined by self-administered questionnaire) was evaluated in a double-blind, placebo-controlled trial in 43 patients with primary and secondary SS. High-viscosity versus low-viscosity xanthan gum-based saliva substitutes were also compared in 33 SS patients. Salivary flow rates (SFR) were determined to examine correlations between the SFR and the subjective efficacy of the saliva substitute. RESULTS: Neither the saliva substitutes nor the placebo was truly effective. Preference for a particular saliva substitute over placebo was equally distributed among the 3 types of substitutes. The SFR of patients who preferred polyacrylic acid-based saliva substitutes was lower than that in patients who preferred the porcine mucin-based substitute (P < 0.05). Patients whose oral dryness was reduced by low-viscoelastic substitutes had a low stimulated SFR ( < 0.20 ml/minute; P < 0.05). CONCLUSION: The optimal properties of a saliva substitute are not the same for all patients with SS, but are dependent on such parameters as the individual SFR. Thus, to determine the best saliva substitute for a particular patient, it is necessary to have the patient try a number of substitutes of different viscoelastic properties.

Rheological properties of commercially available polysaccharides with potential use in saliva substitutes.

The rheological properties of a number of natural and synthetic polysaccharides have been compared with porcine gastric mucin (PGM), a mucin-containing saliva substitute (Saliva Orthana) and with clarified human whole saliva (CHWS). The effects of ionic strength, pH and calcium and fluoride ions on the viscoelastic properties of these polymers have been investigated. Of the polysaccharides tested, xanthan gum and scleroglucan appeared to resemble CHWS most in viscoelastic behavior and may be potential candidates for use in artificial saliva. Both PGM and Saliva Orthana, however, did not show any elastic behavior, whereas a viscosity comparable to human saliva was only observed in highly concentrated solutions. Of the polysaccharides tested, scleroglucan also had mucin-adhesive properties resulting in rheological synergism. This may be the first step in mucoadhesion which may protect underlying oral surfaces in vivo.

[Saliva and saliva substitutes]. / Speeksel en speekselsubstituten.

Saliva is a mixture of secretions of both the large salivary glands (glandula sublingualis submandibularis and parotis) and the minor salivary glands of the palate, tongue, lips and cheeks. The rheological flow of the separate glandular salivas differs remarkably. Sublingual saliva is both more viscous and elastic than the other glandular salivas. The viscosity of parotid saliva, which is of importance during eating (digestion), is equal to the viscosity of water. Utilization of the combination of viscosity and elasticity may improve new saliva substitutes and of frequent applications of saliva substitutes by xerostomia patients may reduce. Besides, the utilization of correct visco-elastic properties of saliva substitutes may lead to better moistening and protective properties.

Shear rate dependent viscoelastic behavior of human glandular salivas.

Rheological properties of unstimulated human whole saliva (CHWS) and human glandular salivas (parotid, submandibular, sublingual, and palatal) of 7 healthy persons were investigated. The viscosity eta' and elasticity eta" of these salivas were measured as a function of oscillating shear rate gamma on an oscillating capillary viscoelasticity analyzer (Vilastic 3). Viscosity eta' and elasticity eta" of total and glandular salivas decreased in the following order: SL > Pal approximately CHWS approximately SM > Par. Rheological behavior of submandibular, palatal and sublingual saliva displayed a comparable pattern, although sublingual saliva showed significantly higher absolute values. The difference in viscoelasticity between submandibular and sublingual saliva was not due to differences in mucin concentration between SM and SL saliva. Flow curves of a range of SL saliva dilutions and flow curves of concentrated SM saliva showed that sublingual saliva was intrinsically more elastic than submandibular saliva.