Socio-economic status and access to fluoridated water in Queensland: an ecological data linkage study.

OBJECTIVES: To investigate the relationship between access to fluoridated drinking water and area-level socio-economic status in Queensland. STUDY DESIGN: Ecological, geospatial data linkage study. SETTING: Queensland, by statistical area level 2 (SA2), 2021. MAIN OUTCOME MEASURES: Proportion of SA2s and of residents with access to fluoridated drinking water (natural or supplemented); relationship at SA2 level between access to fluoridated water and socio-economic status (Index of Relative Socio-economic Advantage and Disadvantage, IRSAD; Index of Economic Resources, IER). RESULTS: In 2021, an estimated 4 050 168 people (79.4% of the population) and 397 SA2 regions (72.7%) in Queensland had access to fluoridated water. Access was concentrated in the southeastern corner of the state. After adjusting for SA2 population, log area, and population density, the likelihood of access to fluoridated drinking water almost doubled for each 100-rank increase in IRSAD (adjusted odds ratio [aOR], 1.93; 95% confidence interval [CI], 1.59-2.36) or IER (aOR, 1.77; 95% CI, 1.50-2.11). CONCLUSIONS: The 2012 decision to devolve responsibility for water fluoridation decisions and funding from the Queensland government to local councils means that residents in lower socio-economic areas are less likely to have access to fluoridated water than those in more advantaged areas, exacerbating their already greater risk of dental disease. Queensland water fluoridation policy should be revised so that all residents can benefit from this evidence-based public health intervention for reducing the prevalence of dental caries.

Parental recall bias in observational studies: Child dental service use.

BACKGROUND: Parents are frequently used to recall events of exposure and disease as a proxy for their children in observational health research. AIM: To assess the validity of parental recall of children's utilisation of dental services. DESIGN: Parents in the Longitudinal Study of Australian Children (LSAC) were asked to recall their children's overall dental service use in the previous year and whether the study child had received fillings or extractions due to decay in the previous 2 years. True positives were identified through parents who correctly recalled child dental service utilisation with actual dental treatment received in a linked dataset. RESULTS: Of the 10 090 participants from the LSAC, 1290 study children had linked dental treatment data eligible for inclusion in this study. The absolute true-positive rate for parental recall of dental service use in the previous year was 82% (n = 1263). Overall true-positive percentages were lower for recall of fillings (40%) and extractions (7%) in the previous 2 years. Increasing number of recall days was associated with the rate of true-positive recall adjusted for other factors in all three models. CONCLUSION: Accuracy of parental recall for tooth fillings and extractions was low. The use of parental recall for specific dental treatment over a time span of 2 years in further research is not recommended.

Geographic patterns of dental service use in the Child Dental Benefits Schedule: 6 years of claims in the Longitudinal Study of Australian Children.

ISSUE ADDRESSED: This article explores the geographic patterns of claims within the Australian Government's Child Dental Benefits Schedule (CDBS). BACKGROUND: The CDBS is a means-tested schedule implemented in 2014 to improve access to dental services for children. Under the schedule, eligible children receive funding to subsidise dental services. METHODS: This study used data from the Longitudinal Study of Australian Children and linked data from the Medicare universal healthcare system, to examine dental service use amongst a subset of children aged 10 and 14 years. Dental service items were classified using Two-step Cluster Analysis, and appointments were analysed using multinomial logistic regression. Geographic characteristics were included as predictor variables. RESULTS: The study found that the majority of dental appointments were non-operative (70.7%, n = 5808), with diagnostic, radiographic, and preventive items being the most common. There were slightly higher proportions of operative appointments (fillings and extractions) compared with non-operative appointments in remote and very remote areas, low socio-economic areas, and Queensland and Northern Territory. Cluster analysis identified eight groups of non-operative appointments and four groups of operative appointments. New South Wales had a higher proportion of 'prophylactic IV' appointments than any other State and Territory, which included debridement and topical fluoride services. CONCLUSION: Cluster analysis identified distinct groups of non-operative and operative appointments, each with unique characteristics. The distribution of appointments varied by State/Territory and region. SO WHAT: Further research and interventions are needed to ensure equitable access to services and a shift to preventive care for disadvantaged populations of Australian children. Exploring alternative funding models that support clinically relevant claims, rather than maximising financial benefits such as time-based renumeration models should be explored.

Dynamic dentin: A quantitative microscopic assessment of age and spatial changes to matrix architecture, peritubular dentin, and collagens types I and III.

To investigate age and site-related changes to human dentin collagen, sound human teeth collected from donors aged 13-29 (young) and 50-74 (aged) years (n = 9/group) were cut to shallow and deep sites. Dentin collagen orientation and fibril bundling was investigated using the Picrosirius Red (PSR) stain observed under cross-polarized light microscopy (Pol), and collagen distribution was investigated using Confocal Laser Scanning Microscopy (CLSM). Collagen types III to I distribution in peritubular dentin (PTD) was revealed using Herovici stain and brightfield microscopy. Image analysis software and linear mixed modelling quantified outcomes. In situ dentin collagen was observed using Xenon Plasma Focussed Ion Beam Scanning Electron Microscopy (Xe PFIB-SEM). The PSR-Pol analysis revealed less coherently aligned and more bundled collagen fibrils in aged dentin (P = 0.005). Deep inner dentin collagen in both groups were less coherently aligned with reduced bundling. Regardless of age, CLSM showed collagen distribution remained stable; and more collagen type III was detectable in PTD located in inner dentin (Young: P = 0.006; Aged: P = 0.008). Observations following Xe PFIB-SEM cross-sectioning showed apatite-like deposits surrounding large intratubular collagen fibers, and evidence of smaller intertubular dentin collagen fibrils in situ. In conclusion, aging changes collagen network architecture, but not distribution or content.

Effect of dentine site on resin and cement adaptation tested using X-ray and electron microscopy to evaluate bond durability and adhesive interfaces.

Glass ionomer (GI) cements and self-etch (SE) or universal adhesives after etching (ER) adapt variably with dentine. Dentine characteristics vary with depth (deep/shallow), location (central/peripheral), and microscopic site (intertubular/peritubular). To directly compare adhesion to dentine, non-destructive imaging and testing are required. Here, GI, ER, and SE adapted at different dentine depths, locations, and sites were investigated using micro-CT, xenon plasma focused ion beam scanning electron microscopy (Xe PFIB-SEM), and energy dispersive X-ray spectroscopy (EDS). Extracted molars were prepared to deep or shallow slices and treated with the three adhesives. Micro-CT was used to compare changes to air volume gaps, following thermocycling, and statistically analysed using a quantile regression model and Fisher's exact test. The three adhesives performed similarly across dentine depths and locations, yet no change or overall increases and decreases in gaps at all dentine depths and locations were measured. The Xe PFIB-SEM-milled dentine-adhesive interfaces facilitated high-resolution characterization, and element profiling revealed variations across the tooth-material interfaces. Dentine depth and location had no impact on adhesive durability, although microscopic differences were observed. Here we demonstrate how micro-CT and Xe PFIB-SEM can be used to compare variable dental materials without complex multi-stage specimen preparation to minimize artefacts.

Understanding the well-being of dentistry students.

INTRODUCTION: The transactional model of stress is a framework describing the process for coping with stressful events as a relationship between the individual person and environment. This study aimed to investigate the associations between personality, learning environment and experiences of mental health for a cohort of Australian dentistry students. MATERIALS AND METHODS: Students were invited in 2017 to complete an online questionnaire including the Depression, Anxiety and Stress Scale (DASS-21), The Dundee Ready Education Environment Measure (DREEM) and Cloninger's Temperament and Character Inventory (TCIR-140). Students were followed-up one year later, and generalised estimating equations were used. RESULTS: A total of 219 (response 73.5%) students participated in the study. Two personality profiles of dentistry students were identified. Group 1 were significantly higher in the traits persistence, self-directedness, cooperativeness and reward dependence, whereas Group 2 were significantly higher in harm avoidance. Students with Group 2 personality had a 3.12 (CI:1.72-5.65) increased odds of depression compared to Group 1 students. Compared to students with positive perceptions of the learning environment, students with negative perceptions had increased odds of stress (3.48, CI: 1.85-6.53), depression (2.71, CI: 1.57- 4.65) and anxiety (2.59, CI: 1.56-4.28). CONCLUSION: Students with personalities high in levels of self-directedness, cooperativeness and persistence and low in harm avoidance, as found in Group 1, demonstrate high levels of general well-being. Positive perceptions of the dentistry learning environment were found to be an important influence on students stress. This study highlighted a number of factors important to student well-being and provides direction for further investigation of interventions aimed at enhancing student well-being.

A comparison of 2- and 3-dimensional mandibular superimposition techniques against Björk's structural superimposition method.

INTRODUCTION: The purpose of this research was to compare mandibular growth rotation relative to the cranial base in different vertical facial patterns on the basis of multiple 2-dimensional (2D) and 3-dimensional (3D) superimposition methods. METHODS: Cone-beam computed tomography (CBCT) images taken at a mean interval of 54.8 ± 16.8 months were assessed from a sample of 70 growing patients. Three mandibular superimposition methods were compared against Björk's structural method: (1) a 2D landmark method (2D-M1), (2) a voxel-based 3D method based on a previously reported method (3D-M1), and (3) a voxel-based 3D method incorporating symphyseal structures as references (3D-M2). After superimposition, the relative change in cranial base lines as depicted in sagittal views were measured for true mandibular rotation. Agreement between methods was assessed with Lin's concordance correlation coefficient, Bland-Altman's limits of agreement, and the Bradley-Blackwood test. RESULTS: Lin's concordance correlation coefficients ranged between 0.924 for the 2D-M1 method, 0.695 for the 3D-M1 method, and 0.965 for the 3D-M2 method. Bland-Altman limits of agreement were wide for all but the 3D-M2 method. Finally, the Bradley-Blackwood test of equality of means and variances was significant in all except the 3D-M2 method. CONCLUSIONS: For time intervals between CBCT volume acquisitions >3 years, the use of the 2D-M1 and 3D-M1 methods is not recommended. There was a high concordance between the 3D-M2 method and Björk's structural method when assessing mandibular growth rotation using relative changes in cranial base lines. The high concordance was displayed across all vertical facial types and for all time differences between first and second CBCT data acquisitions.

The prevalence and burden of recurrent headache in Australian adolescents: findings from the longitudinal study of Australian children.

BACKGROUND: Headache disorders are highly prevalent worldwide, but not well investigated in adolescents. Few studies have included representative nationwide samples. This study aimed to present the prevalence and burden of recurrent headache in Australian adolescents. METHODS: The prevalence of recurrent headache, headache characteristics (severity and frequency) and burden on health-related quality of life in Australian children aged 10-17 years were presented, using nationally representative data from the Longitudinal Study of Australian children (LSAC). The LSAC, commencing in 2004, collects data every 2 years from a sample of Australian children of two different age cohorts: B 'baby' cohort, aged 0-1 years and K 'kindergarten' cohort, aged 4-5 years at the commencement of the study. Face-to-face interviews and self-complete questionnaires have been conducted with the study child and parents of the study child (carer-reported data) at each data collection wave, with seven waves of data available at the time of the current study. Wave 7 of the LSAC was conducted in 2016, with B cohort children aged 12-13 years and K cohort children aged 16-17 years. For the current study, data were accessed for four out of seven waves of available data (Wave 4-7) and presented cross-sectionally for the two cohorts of Australian children, for the included age groups (10-11 years, 12-13 years, 14-15 years and 16-17 years). All available carer-reported questionnaire data pertaining to headache prevalence, severity and frequency, general health and health-related quality of life, for the two cohorts, were included in the study, and presented for male and female adolescents. Carer-reported general health status of the study child and health-related quality of life scores, using the parent proxy-report of the Paediatric Quality of Life Inventory™ 4.0, were compared for male and female adolescents with recurrent headache and compared with a healthy group. Finally, health-related quality of life scores were compared based on headache frequency and severity. RESULTS: The LSAC study initially recruited 10,090 Australian children (B cohort n = 5107, K cohort n = 4983), and 64.1% of the initial sample responded at wave 7. Attrition rates across the included waves ranged from 26.3% to 33.8% (wave 6 and 7) for the B cohort, and 16.3% to 38.0% (wave 4-7) for the K cohort. Recurrent headache was more common in females, increasing from 6.6% in 10-11 years old females to 13.2% in 16-17 years old females. The prevalence of headache in males ranged from 4.3% to 6.4% across the age groups. Health-related quality of life scores were lower for all functional domains in adolescents with recurrent headache, for both sexes. Headache frequency, but not severity, was significantly associated with lower health-related quality of life scores, in both males and females. CONCLUSIONS: Recurrent headache was common among Australian adolescents and increased in prevalence for females, across the age groups. Frequent recurrent headache is burdensome for both male and female adolescents. This study provides information regarding the prevalence and burden of recurrent headache in the adolescent population based on findings from the Longitudinal Study of Australian Children.

Intervention To Stop Transmission of Imported Pneumonic Plague - Uganda, 2019.

Plague, an acute zoonosis caused by Yersinia pestis, is endemic in the West Nile region of northwestern Uganda and neighboring northeastern Democratic Republic of the Congo (DRC) (1-4). The illness manifests in multiple clinical forms, including bubonic and pneumonic plague. Pneumonic plague is rare, rapidly fatal, and transmissible from person to person via respiratory droplets. On March 4, 2019, a patient with suspected pneumonic plague was hospitalized in West Nile, Uganda, 4 days after caring for her sister, who had come to Uganda from DRC and died shortly thereafter, and 2 days after area officials received a message from a clinic in DRC warning of possible plague. The West Nile-based Uganda Virus Research Institute (UVRI) plague program, together with local health officials, commenced a multipronged response to suspected person-to-person transmission of pneumonic plague, including contact tracing, prophylaxis, and education. Plague was laboratory-confirmed, and no additional transmission occurred in Uganda. This event transpired in the context of heightened awareness of cross-border disease spread caused by ongoing Ebola virus disease transmission in DRC, approximately 400 km to the south. Building expertise in areas of plague endemicity can provide the rapid detection and effective response needed to mitigate epidemic spread and minimize mortality. Cross-border agreements can improve ability to respond effectively.

Virtual Reality as a novel educational tool in pre-clinical paediatric dentistry training: Students' perceptions.

BACKGROUND: Dental students are required to demonstrate competency by pre-clinical simulated practice before performing invasive clinical procedures on patients. The Moog Simodont® Dental Trainer provides a virtual reality-based dental simulation environment for training students. AIM: This cross-sectional questionnaire-based study compared students' perception of the pre-clinical paediatric dentistry training gained in Simodont® and conventional simulation environment. DESIGN: The dental students who completed pulpotomies and stainless steel crowns (SSCs) training in Simodont® and conventional pre-clinical simulation laboratory were invited to complete a questionnaire on their experience in both environments. The percentages for the distribution of responses to statements about training modality were tabulated, and intra-participant comparisons were used to measure student preference for either Simodont® or conventional simulation training. RESULTS: One hundred students completed the survey. Fifty-one per cent of students agreed that using Simodont® assisted their learning, and 56% felt Simodont® training facilitated their understanding of paediatric dentistry tasks. Generally, participants felt more comfortable with simulation training than Simodont® for both practical exercises. Eighty-eight per cent of the participants disagreed that Simodont® should replace conventional simulation. CONCLUSIONS: The study suggests that Simodont® could be used as an adjunct in training dental students for pre-clinical paediatric dentistry restorative exercises.

Predicting a beneficial response to motor control training in patients with low back pain: a longitudinal cohort study.

PURPOSE: Exercise therapy such as motor control training (MCT) has been shown to reduce pain and disability in people with low back pain (LBP). It is unknown which patients are most likely to benefit. This longitudinal cohort study aimed to: (1) retrospectively examine records from a large cohort of patients who received MCT treatment, (2) identify potentially important predictors of response to MCT and (3) test the predictors on an independent (split) sample derived from the original cohort of patients, using one group to identify the predictors and the other to test them. METHODS: The response of 775 patients with LBP to MCT was classified as 'improved' or 'not improved' based on self-reported change in pain levels and symptoms. Measures were examined for associations with improvement and entered into a logistic regression model to classify patients as low, medium or high benefits of improvement with MCT. The model was tested on an independent sample. RESULTS: A positive response was seen in patients with: no evidence of scoliosis [OR = 4.0, 95% CI (1.7, 9.6)], LBP without associated groin pain [OR = 2.2, 95% CI (1.0, 5.0)], LBP which was chronic recurrent in nature [OR = 3.1, 95% CI (1.8, 5.3)] and poor results on muscle testing of the multifidus muscle [OR = 2.0, 95% CI (1.1, 3.7)]. Testing on an independent sample confirmed that patients could be classified into benefit groups. CONCLUSIONS: This study provides a first step towards assisting clinicians to select patients most likely to respond to MCT. These slides can be retrieved under Electronic Supplementary Material.

Detection of Tickborne Relapsing Fever Spirochete, Austin, Texas, USA.

In March 2017, a patient became febrile within 4 days after visiting a rustic conference center in Austin, Texas, USA, where Austin Public Health suspected an outbreak of tickborne relapsing fever a month earlier. Evaluation of a patient blood smear and molecular diagnostic assays identified Borrelia turicatae as the causative agent. We could not gain access to the property to collect ticks. Thus, we focused efforts at a nearby public park, <1 mile from the suspected exposure site. We trapped Ornithodoros turicata ticks from 2 locations in the park, and laboratory evaluation resulted in cultivation of 3 B. turicatae isolates. Multilocus sequencing of 3 chromosomal loci (flaB, rrs, and gyrB) indicated that the isolates were identical to those of B. turicatae 91E135 (a tick isolate) and BTE5EL (a human isolate). We identified the endemicity of O. turicata ticks and likely emergence of B. turicatae in this city.

Successful Treatment of Human Plague with Oral Ciprofloxacin.

The US Food and Drug Administration recently approved ciprofloxacin for treatment of plague (Yersina pestis infection) based on animal studies. Published evidence of efficacy in humans is sparse. We report 5 cases of culture-confirmed human plague treated successfully with oral ciprofloxacin, including 1 case of pneumonic plague.

Patterns of Human Plague in Uganda, 2008-2016.

Plague is a highly virulent fleaborne zoonosis that occurs throughout many parts of the world; most suspected human cases are reported from resource-poor settings in sub-Saharan Africa. During 2008-2016, a combination of active surveillance and laboratory testing in the plague-endemic West Nile region of Uganda yielded 255 suspected human plague cases; approximately one third were laboratory confirmed by bacterial culture or serology. Although the mortality rate was 7% among suspected cases, it was 26% among persons with laboratory-confirmed plague. Reports of an unusual number of dead rats in a patient's village around the time of illness onset was significantly associated with laboratory confirmation of plague. This descriptive summary of human plague in Uganda highlights the episodic nature of the disease, as well as the potential that, even in endemic areas, illnesses of other etiologies might be being mistaken for plague.

Evaluation of bioMérieux's Dissociated Vidas Lyme IgM II and IgG II as a First-Tier Diagnostic Assay for Lyme Disease.

The recommended laboratory diagnostic approach for Lyme disease is a standard two-tiered testing (STTT) algorithm where the first tier is typically an enzyme immunoassay (EIA) that if positive or equivocal is reflexed to Western immunoblotting as the second tier. bioMérieux manufactures one of the most commonly used first-tier EIAs in the United States, the combined IgM/IgG Vidas test (LYT). Recently, bioMérieux launched its dissociated first-tier tests, the Vidas Lyme IgM II (LYM) and IgG II (LYG) EIAs, which use purified recombinant test antigens and a different algorithm than STTT. The dissociated LYM/LYG EIAs were evaluated against the combined LYT EIA using samples from 471 well-characterized Lyme patients and controls. Statistical analyses were conducted to assess the performance of these EIAs as first-tier tests and when used in two-tiered algorithms, including a modified two-tiered testing (MTTT) approach where the second-tier test was a C6 EIA. Similar sensitivities and specificities were obtained for the two testing strategies (LYT versus LYM/LYG) when used as first-tier tests (sensitivity, 83 to 85%; specificity, 85 to 88%) with an observed agreement of 80%. Sensitivities of 68 to 69% and 76 to 77% and specificities of 97% and 98 to 99% resulted when the two EIA strategies were followed by Western immunoblotting and when used in an MTTT, respectively. The MTTT approach resulted in significantly higher sensitivities than did STTT. Overall, the LYM/LYG EIAs performed equivalently to the LYT EIA in test-to-test comparisons or as first-tier assays in STTT or MTTT with few exceptions.

Ebola Virus Disease Diagnostics, Sierra Leone: Analysis of Real-time Reverse Transcription-Polymerase Chain Reaction Values for Clinical Blood and Oral Swab Specimens.

During the Ebola virus outbreak of 2013-2016, the Viral Special Pathogens Branch field laboratory in Sierra Leone tested approximately 26 000 specimens between August 2014 and October 2015. Analysis of the B2M endogenous control Ct values showed its utility in monitoring specimen quality, comparing results with different specimen types, and interpretation of results. For live patients, blood is the most sensitive specimen type and oral swabs have little diagnostic utility. However, swabs are highly sensitive for diagnostic testing of corpses.

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples.

The current recommendation for the laboratory confirmation of Lyme disease is serology-based diagnostics. Specifically, a standardized two-tiered testing (STTT) algorithm is applied that utilizes a first-tier immunofluorescence assay or enzyme immunoassay (EIA) that, if the result is positive or equivocal, is followed by second-tier immunoblotting. Despite the standardization and performance achievements, STTT is considered technically complex and subjective, as well as insensitive for early acute infection. These issues have prompted development of novel algorithms and testing platforms. In this study, we evaluated the performance of several commonly used assays for STTT. Several modified two-tiered testing (MTTT) algorithms, including a 2-EIA algorithm and modified criteria for second-tier IgG immunoblots, were also evaluated. All tests were performed on sera from a recently available, well-defined archive of positive- and negative-control patients. Our study demonstrates differences in the results between individual first- and second-tier tests, although the overall agreement of the different STTT algorithms used was strong. In addition, the MTTT algorithm utilizing 2-EIAs was found to be equivalent to all STTT algorithms tested, with agreement ranging from 94 to 97%. The 2-EIA MTTT algorithm slightly enhanced sensitivity in early disease compared to the STTT algorithms evaluated. Furthermore, these data add to the mounting evidence that a 2-EIA-based MTTT algorithm, where immunoblotting is replaced by the C6 EIA, performs as well or better than STTT.

Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus.

Acute febrile illness (AFI) is associated with substantial morbidity and mortality worldwide, yet an etiologic agent is often not identified. Convalescent-phase serology is impractical, blood culture is slow, and many pathogens are fastidious or impossible to cultivate. We developed a real-time PCR-based TaqMan array card (TAC) that can test six to eight samples within 2.5 h from sample to results and can simultaneously detect 26 AFI-associated organisms, including 15 viruses (chikungunya, Crimean-Congo hemorrhagic fever [CCHF] virus, dengue, Ebola virus, Bundibugyo virus, Sudan virus, hantaviruses [Hantaan and Seoul], hepatitis E, Marburg, Nipah virus, o'nyong-nyong virus, Rift Valley fever virus, West Nile virus, and yellow fever virus), 8 bacteria (Bartonella spp., Brucella spp., Coxiella burnetii, Leptospira spp., Rickettsia spp., Salmonella enterica and Salmonella enterica serovar Typhi, and Yersinia pestis), and 3 protozoa (Leishmania spp., Plasmodium spp., and Trypanosoma brucei). Two extrinsic controls (phocine herpesvirus 1 and bacteriophage MS2) were included to ensure extraction and amplification efficiency. Analytical validation was performed on spiked specimens for linearity, intra-assay precision, interassay precision, limit of detection, and specificity. The performance of the card on clinical specimens was evaluated with 1,050 blood samples by comparison to the individual real-time PCR assays, and the TAC exhibited an overall 88% (278/315; 95% confidence interval [CI], 84% to 92%) sensitivity and a 99% (5,261/5,326, 98% to 99%) specificity. This TaqMan array card can be used in field settings as a rapid screen for outbreak investigation or for the surveillance of pathogens, including Ebola virus.

Collection and characterization of samples for establishment of a serum repository for lyme disease diagnostic test development and evaluation.

Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first- and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests.