Clinical Validation of a Colorimetric Loop-Mediated Isothermal Amplification Using a Portable Device for the Rapid Detection of SARS-CoV-2.

Quick and reliable mass testing of infected people is an effective tool for the contingency of SARS-CoV-2. During the COVID-19 pandemic, Point-of-Care (POC) tests using Loop-Mediated Isothermal Amplification (LAMP) arose as a useful diagnostic tool. LAMP tests are a robust and fast alternative to Polymerase Chain Reaction (PCR), and their isothermal property allows easy incorporation into POC platforms. The main drawback of using colorimetric LAMP is the reported short-term stability of the pre-mixed reagents, as well as the relatively high rate of false-positive results. Also, low-magnitude amplification can produce a subtle color change, making it difficult to discern a positive reaction. This paper presents Hilab Molecular, a portable device that uses the Internet of Things and Artificial Intelligence to pre-analyze colorimetric data. In addition, we established manufacturing procedures to increase the stability of colorimetric RT-LAMP tests. We show that ready-to-use reactions can be stored for up to 120 days at -20 °C. Furthermore, we validated both the Hilab Molecular device and the Hilab RT-LAMP test for SARS-CoV-2 using 581 patient samples without any purification steps. We achieved a sensitivity of 92.93% and specificity of 99.42% (samples with CT ≤ 30) when compared to RT-qPCR.

Cost-Effectiveness of Molecular Method Diagnostic for Rapid Detection of Antibiotic-Resistant Bacteria.

OBJECTIVES: This study aimed to perform a cost-effectiveness analysis (CEA) of the molecular diagnostic method (MM) associated with conventional diagnostic method (CM) compared with the CM alone, for the detection of resistant profile in bacteremia, from the perspective of the Brazilian Public Health System, in intensive care units setting. METHODS: The clinical parameters regarding methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Gram-negative bacteria (CRGNB), and vancomycin-resistant Enterococcus spp. (VRE) infections were collected from searches on PubMed, Scopus, and SciELO, using specific keywords. Data on direct medical costs to treat these infections were collected according to Brazilian Public Health System perspective from Brazilian databases, in tables of 2018 to 2019. CEA was performed after building a dynamic model, which was calibrated and validated according to international recommendations. The incremental cost-effectiveness ratio of the MM + CM compared with the CM was calculated using the outcomes "avoided death" and "avoided resistant infections." One-way sensitivity analyses were performed. RESULTS: This CEA demonstrated that the MM + CM was dominant in all scenarios. Estimates showed that for MRSA, CRGNB, and VRE infections, every avoided death would lead to savings of Brazilian real (R$) 4.9 million ($937 301), R$2.2 million ($419 899), and R$1.3 million ($248 919), respectively. The same infections assessed by avoided resistant infections savings were projected to be R$24 964 ($4686), R$40 260 ($7558), and R$23 867 ($4480). CONCLUSIONS: MM leads to cost reduction and increased benefits, optimizing the use of financial resources on the health system in the intensive care unit setting, in bacteremia caused by MRSA, CRGNB, and VRE.

Rapid Serological Tests for SARS-CoV-2: Diagnostic Performance of 4 Commercial Assays.

OBJECTIVE: To assess the diagnostic performance of lateral flow immunochromatographic assays (LFAs) of 4 different manufacturers to identify SARS-CoV-2 antibodies (IgM, IgG, or total), comparing them with the nucleic acid amplification test (NAAT) or the clinical defined test (definite or probable SARS-CoV-2 infection, respectively). METHODS: One hundred nineteen serum samples were randomly selected by convenience and distributed in the following groups: (1) group with SARS-CoV-2 infection (n = 82; RT-qPCR positive [definite, n = 70] and probable [n = 12]); (2) other diseases (n = 27; other viruses identified [n = 8] and SARS of other etiologies [n = 19]); and (3) healthy control group (n = 10). LFAs of 4 manufacturers were compared: MedTest Coronavirus (COVID-19) IgG/IgM (MedLevensohn, Brazil); COVID-19 IgG/IgM ECO Test (Ecodiagnóstica, Brazil); Camtech COVID-19 IgM/IgG Rapid Test Kit (Camtech Diagnostics Pte Ltd, Singapore); and 1-Step COVID-19 Test for total antibodies (Guangzhou Wondfo Biotech Co., China). RESULTS: The 4 tests studied showed high diagnostic performance characteristics for the diagnoses of definite or probable SARS-CoV-2 infection. The best measures were for the Wondfo test: sensitivity (86.59%; 95% CI: 77.26-93.11%), specificity (100%; 90.51-100%), DOR (257; 60-1,008), LR+ (33.43; 4.82-231.85), LR- (0.13; 0.08-0.23), accuracy (90.76%; 84.06-95.29%), and Matthews correlation coefficient (MCC) 0.82. Although considering only the probable SARS-CoV-2 infection (PCR-) cases, all the kits studied showed limited values. CONCLUSION: Our data demonstrate the excellent performance of LFA for the diagnoses of definite or probable SARS-CoV-2 infection. There was substantial heterogeneity in sensitivities of IgM and IgG antibodies among the different kits. LFA tests cannot replace molecular diagnostics but should be used as an additional screening tool.

Charlson comorbidity index scores and in-hospital prognosis of patients with severe acute respiratory infections.

BACKGROUND: Respiratory infections are one of the leading causes of mortality, and comorbid conditions play a significant role in the severity and fatality of these infections. AIMS: We evaluated the Charlson Comorbidity Index (CCI) score and possible predictors of mortality in hospitalised patients with severe acute respiratory infection (SARI), aiming to test if the CCI is a valid in-hospital prognostic indicator. METHODS: Patients older than 14 years, hospitalised from 2010 to 2016 due to SARI by viral infection and who were submitted to respiratory virus testing were included. We assessed comorbidity retrospectively through chart review and calculated four variants of the CCI. RESULTS: Of the 291 patients assessed, 72.8% (n = 212) presented comorbidities, and 24% died (n = 70). The most recurrent comorbidities were chronic pulmonary disease (n = 76/212, 36%) and HIV (n = 50/212, 23.6%). The 1994 age-adjusted CCI predicted in-hospital mortality in SARI patients (P = 0.04), and HIV was associated with in-hospital mortality (P = 0.032). CONCLUSIONS: The comorbidity scores used to assess mortality risk in hospitalised patients with SARI displayed poor results, but HIV infection was considered a marker of severity. However, other factors should be considered in order to compose a score system that allows us to specifically assess the risk of mortality in patients with SARI.

Matching the Dimensions of Currently Available Instruments with the Apical Diameters of Mandibular Molar Mesial Root Canals Obtained by Micro-computed Tomography.

INTRODUCTION: This micro-computed tomographic study investigated the original anatomic diameters of the apical portion of mandibular molar mesial canals and matched them to the dimensions of instruments that are currently available and commonly used for apical preparation. METHODS: One hundred eight mandibular molar mesial roots with Vertucci type IV configuration were scanned by micro-computed tomographic imaging, and the largest anatomic diameter of both mesial canals at 1, 2, 3, and 4 mm short of the apical foramen was measured. Canal diameters were compared with the dimensions of 10 endodontic instruments for simulation of preparation 1 mm short of the apical foramen. Accordingly, the instruments with a diameter larger than the largest canal diameter were regarded as having the potential to prepare 100% of the canal walls at each specific point or over the 4-mm apical segment. RESULTS: At 1 and 2 mm short of the apical foramen, a 45/.02 instrument had a larger diameter than the mesiobuccal canal diameter in 73% and 55% of the teeth, respectively. Corresponding figures for the mesiolingual canal were 65% and 55%. When the entire 4-mm apical segment was considered, a 40/.06 instrument had better results, being larger than the apical canal in about 20% of the specimens. Overall, 78% of the apical canals would not be completely prepared by any instrument. Based on the mean (median) anatomic diameters, the adequate instrument dimensions would be 40/.10 (40/.08) for mesiobuccal canals and 45/.08 (40/.09) for mesiolingual canals. CONCLUSIONS: The dimensions of the available instruments are not compatible with complete apical preparation in the majority of cases. The ideal instrument size/taper to include all the canal walls in apical preparation is too large and may not be safe for clinical use.

Impact and seasonality of human rhinovirus infection in hospitalized patients for two consecutive years / Impacto e sazonalidade da infecção por rinovírus humano em pacientes internados por dois anos consecutivos

Abstract Objectives: To report epidemiological features, clinical characteristics, and outcomes of human rhinovirus (HRV) infections in comparison with other community acquired respiratory virus (CRV) infections in patients hospitalized for two consecutive years. Methods: This was a cross-sectional study. Clinical, epidemiological, and laboratory data of patients hospitalized with acute respiratory syndrome in a tertiary care hospital from 2012 to 2013 were reviewed. Results: HRV was the most common CRV observed (36%, 162/444) and was present in the majority of viral co-detections (69%, 88/128), mainly in association with human enterovirus (45%). Most HRV-infected patients were younger than 2 years (57%). Overall, patients infected with HRV had a lower frequency of severe acute respiratory infection than those infected with other CRVs (60% and 84%, respectively, p = 0.006), but had more comorbidities (40% and 27%, respectively; p = 0.043). However, in the adjusted analysis this association was not significant. The mortality rate within the HRV group was 3%. Detection of HRV was more prevalent during autumn and winter, with a moderately negative correlation between viral infection frequency and temperature (r = −0.636, p < 0.001) but no correlation with rainfall (r = −0.036, p = 0.866). Conclusion: HRV is usually detected in hospitalized children with respiratory infections and is often present in viral co-detections. Comorbidities are closely associated with HRV infections. These infections show seasonal variation, with predominance during colder seasons.

Resumo Objetivos: Relatar as características epidemiológicas, as características clínicas e os resultados das infecções por rinovírus humano (RVH) em comparação a outras infecções por vírus respiratórios adquiridos na comunidade (VRCs) em pacientes internados por dois anos consecutivos. Métodos: Este foi um estudo transversal. Foram revisados os dados clínicos, epidemiológicos e laboratoriais de pacientes internados com síndrome respiratória aguda em um hospital terciário de 2012 a 2013. Resultados: O RVH foi o VRC mais comum observado (36%, 162/444) e esteve presente na maior parte das codetecções virais (69%, 88/128), principalmente em associação ao enterovírus humano (45%). A maioria dos pacientes infectados por RVH possuía menos de 2 anos (57%). De modo geral, os pacientes com RVH apresentaram uma menor frequência de infecção respiratória aguda grave que os pacientes infectados por outros VRCs (60% e 84%, respectivamente, p = 0,006), porém mais comorbidades (40% e 27%, respectivamente; p = 0,043). Contudo, em uma análise ajustada, essa associação não foi significativa. A taxa de mortalidade no grupo RVH foi 3%. A detecção de RVH foi mais prevalente durante o outono e inverno, com uma correlação negativa moderada entre a frequência de infecção viral e a temperatura (r = -0,636, p < 0,001), porém nenhuma correlação com a precipitação (r = −0,036, p = 0,866). Conclusão: O RVH é normalmente detectado em crianças internadas com infecções respiratórias e normalmente está presente em codetecções virais. As comorbidades estão estreitamente associadas a infecções por RVH. Essas infecçõesmostram variação sazonal, com predominância durante as estações mais frias.

Flavivirus cross-reactivity in serological tests and Guillain-Barré syndrome in a hematopoietic stem cell transplant patient: A case report.

Serological diagnosis of flavivirus infection is a challenge, particularly in the context of a disease associated with immune response enhancement in a transplant patient, where aspects such as previous flavivirus infections may be involved with the outcome. We report a case of a pediatric patient who developed Guillain-Barré syndrome (GBS) after matched-unrelated hematopoietic stem cell transplantation (HSCT). The patient lives in a Brazilian region that is experiencing an epidemic of Zika virus (ZIKV) and dengue virus (DENV). Because an increasing number of cases of GBS, likely triggered by ZIKV infection, are being reported in Brazil, samples from the patient were tested for both ZIKV and DENV infection. Serological assays strongly suggested a recent ZIKV infection, although infection by DENV or co-infection with both viruses cannot be ruled out. The presence of anti-DENV immunoglobulin-G in donor serum led to the hypothesis that antibodies from the donor could have enhanced the severity of the ZIKV infection. This hypothesis is in agreement with the recent findings that DENV sero-cross-reactivity drives antibody-dependent enhancement of ZIKV infection. These findings highlight the need for discussion of the indication to perform previous flavivirus tests in HSCT donors, especially in areas where ZIKV and other flaviviruses co-circulate.

Impact and seasonality of human rhinovirus infection in hospitalized patients for two consecutive years.

OBJECTIVES: To report epidemiological features, clinical characteristics, and outcomes of human rhinovirus (HRV) infections in comparison with other community acquired respiratory virus (CRV) infections in patients hospitalized for two consecutive years. METHODS: This was a cross-sectional study. Clinical, epidemiological, and laboratory data of patients hospitalized with acute respiratory syndrome in a tertiary care hospital from 2012 to 2013 were reviewed. RESULTS: HRV was the most common CRV observed (36%, 162/444) and was present in the majority of viral co-detections (69%, 88/128), mainly in association with human enterovirus (45%). Most HRV-infected patients were younger than 2 years (57%). Overall, patients infected with HRV had a lower frequency of severe acute respiratory infection than those infected with other CRVs (60% and 84%, respectively, p=0.006), but had more comorbidities (40% and 27%, respectively; p=0.043). However, in the adjusted analysis this association was not significant. The mortality rate within the HRV group was 3%. Detection of HRV was more prevalent during autumn and winter, with a moderately negative correlation between viral infection frequency and temperature (r=-0.636, p<0.001) but no correlation with rainfall (r=-0.036, p=0.866). CONCLUSION: HRV is usually detected in hospitalized children with respiratory infections and is often present in viral co-detections. Comorbidities are closely associated with HRV infections. These infections show seasonal variation, with predominance during colder seasons.

Correlative bacteriologic and micro-computed tomographic analysis of mandibular molar mesial canals prepared by self-adjusting file, reciproc, and twisted file systems.

INTRODUCTION: This ex vivo study evaluated the disinfecting and shaping ability of 3 protocols used in the preparation of mesial root canals of mandibular molars by means of correlative bacteriologic and micro-computed tomographic (µµCT) analysis. METHODS: The mesial canals of extracted mandibular molars were contaminated with Enterococcus faecalis for 30 days and assigned to 3 groups based on their anatomic configuration as determined by µCT analysis according to the preparation technique (Self-Adjusting File [ReDent-Nova, Ra'anana, Israel], Reciproc [VDW, Munich, Germany], and Twisted File [SybronEndo, Orange, CA]). In all groups, 2.5% NaOCl was the irrigant. Canal samples were taken before (S1) and after instrumentation (S2), and bacterial quantification was performed using culture. Next, mesial roots were subjected to additional µCT analysis in order to evaluate shaping of the canals. RESULTS: All instrumentation protocols promoted a highly significant intracanal bacterial reduction (P < .001). Intergroup quantitative and qualitative comparisons disclosed no significant differences between groups (P > .05). As for shaping, no statistical difference was observed between the techniques regarding the mean percentage of volume increase, the surface area increase, the unprepared surface area, and the relative unprepared surface area (P > .05). Correlative analysis showed no statistically significant relationship between bacterial reduction and the mean percentage increase of the analyzed parameters (P > .05). CONCLUSIONS: The 3 instrumentation systems have similar disinfecting and shaping performance in the preparation of mesial canals of mandibular molars.

Time-dependent antibacterial effects of the self-adjusting file used with two sodium hypochlorite concentrations.

INTRODUCTION: This in vitro study evaluated the time-dependent ability of the self-adjusting file (SAF) used with either of two NaOCl concentrations to reduce bacterial populations in long oval root canals. METHODS: Oval-shaped canals from extracted teeth were infected with Enterococcus faecalis for 30 days and subjected to preparation with the SAF system using continuous irrigation with either 2.5% NaOCl (n = 11) or 6% NaOCl (n = 11). Bacteriological samples were taken before preparation (baseline) and after 2, 4, and 6 minutes of SAF operation. RESULTS: Intragroup quantitative analyses revealed that the SAF operating for 2, 4, or 6 minutes yielded a highly significant bacterial reduction when compared with counts at baseline (P < .001) irrespective of the NaOCl concentration. No significant differences were found for intragroup analyses comparing the three time periods (P > .05). Likewise, intergroup quantitative analysis showed no significant difference for the SAF with 2.5% NaOCl or 6% NaOCl during 2, 4, or 6 minutes (P > .05). The incidence of positive cultures in the 2.5% NaOCl group was 6 of 11, 5 of 11, and 2 of 11 after 2, 4, and 6 minutes, respectively. Corresponding figures in the 6% NaOCl group were 4 of 11, 2 of 11, and 1 of 11. When all samples were gathered together, overall findings revealed that using the SAF for 6 minutes significantly reduced both the bacterial counts (P = .02) and the incidence of positive cultures (P = .04) when compared with 2 minutes. CONCLUSIONS: Findings revealed that the SAF promoted a significant reduction in bacterial populations even after only 2 minutes regardless of the NaOCl concentration. The most impressive results were obtained after a 6-minute operation.

Disinfecting oval-shaped root canals: effectiveness of different supplementary approaches.

INTRODUCTION: This study compared the ability of different approaches to supplement the antibacterial effects of chemomechanical preparation in oval-shaped root canals. METHODS: Long oval canals from extracted teeth infected with Enterococcus faecalis (ATCC 29212) were chemomechanically prepared up to a 40/04 rotary BioRaCe instrument using 2.5% NaOCl irrigation and then subjected to two supplementary protocols. In the passive ultrasonic irrigation (PUI)/chlorhexidine (CHX) group, canals were subjected to PUI for the activation of NaOCl followed by a final rinse with 0.2% CHX digluconate solution. In the Hedström group, canals received additional Hedström filing directed towards the buccal and lingual canal recesses. Bacteriological samples were taken before and after preparation, after PUI or Hedström instrumentation, and after CHX final rinsing. RESULTS: Chemomechanical preparation and the supplementary steps promoted a highly significant bacterial reduction (P < .001). Quantitative (reduction in levels) and qualitative (frequency of negative cultures) analyses showed that PUI alone or Hedström filing did not significantly increase bacterial reduction (P > .05). Further rinsing with CHX also failed to significantly increase bacterial elimination when compared with post-PUI samples. However, the cumulative antibacterial effects of PUI and CHX final rinse were effective in significantly reducing bacterial counts to levels below those achieved after preparation (P = .03). This combined PUI/CHX approach also resulted in a significant increase in the incidence of negative cultures (P = .04). CONCLUSIONS: Findings suggest that there may be a benefit of using the PUI for the activation of NaOCl followed by a final rinse with CHX as supplementary steps in the treatment of infected oval-shaped root canals.

Ability of chemomechanical preparation with either rotary instruments or self-adjusting file to disinfect oval-shaped root canals.

INTRODUCTION: Oval-shaped root canals might represent a great challenge for proper disinfection. This study compared the capability of a newly developed instrument, the self-adjusting file (SAF), and rotary nickel-titanium (NiTi) instrumentation to eliminate Enterococcus faecalis populations from long oval root canals of extracted human teeth. As a secondary purpose, the ability of a modification in sampling technique to recover bacteria lodged in recesses of oval canals was evaluated. METHODS: Long oval canals from mandibular incisors and maxillary second premolars were infected with E. faecalis (ATCC 29212) for 30 days and then randomly distributed into 2 experimental groups. In group 1, canals were prepared up to a 40/04 rotary BioRaCe instrument by using irrigation with NaviTip needles; in group 2, canals were prepared by using the SAF system with continuous irrigation. NaOCl and ethylenediaminetetraacetic acid were used as irrigants. Bacteriologic samples were taken before (S1) and after preparation (S2a and S2b). RESULTS: Reduction in the bacterial populations was highly significant in both groups (P < .001). Preparation of long oval canals with the SAF was significantly more effective than rotary NiTi instrumentation in reducing intracanal E. faecalis counts (P = .01). Frequency of positive cultures in S2 samples was 11 of 20 (55%) for rotary instrumentation and 4 of 20 (20%) for SAF instrumentation (P = .048). S2b samples (modified method) yielded more positive samples than S2a (12/40 vs 5/40), but this difference reached no statistical significance (P > .05). CONCLUSIONS: The SAF system was significantly more effective than rotary NiTi instrumentation used with syringe/needle irrigation in disinfecting long oval root canals in vitro. A modified sampling technique might be necessary for oval canals.