Commercial Test Research Articles

Immune response and laboratory markers in the spectrum of severity of COVID-19

BACKGROUND: Serum antibodies to SARS-CoV-2, when measured early after disease onset, may add value to the diagnosis of COVID-19. AIM: To examine the levels of serum antibodies to SARS-CoV-2 and laboratory blood parameters in hospitalized patients with COVID-19 of varying severity. MATERIALS AND METHODS: In this retrospective cohort study, we examined laboratory markers of inflammation in patients with acute COVID-19 during the 1st week of hospitalization. The levels of serum antibodies to SARS-CoV-2 were studied using commercial test systems. RESULTS: In 47% of hospitalized patients with COVID-19, during the first week of hospital stay, IgM and IgG antibodies to SARS-CoV-2 were detected, both in the case of a positive and negative PCR test. An average positive cor- relation of detected IgM and IgG with antibodies to the receptor-binding site of the S-protein of the SARS-CoV-2 virus is shown. In total, IgM and IgG antibodies to SARS-CoV-2 were most often detected in patients with a favorable course of the disease. Laboratory parameters in patients with moderate and severe COVID-19 were characterized by asignificant increase in the level of serum C-reactive protein, an increase in the neutrophil-leukocyte ratio and fibrinogen level, in comparison with data from patients with a mild course of the disease. In mild cases of infection, a moderately negative correlation was revealed between the levels of antibodies to SARS-CoV-2 and NLR. CONCLUSIONS: Detection of antibodies to SARS-CoV-2 in the early stages of hospitalization may be a predictor of a favorable outcome of the disease and serve as an additional criterion for the diagnosis of COVID-19 along with PCR analysis.

Lateral Flow Assay Biotesting by Utilizing Plasmonic Nanoparticles Made of Inexpensive Metals─Replacing Colloidal Gold.

Nanoparticles (NPs) can be conjugated with diverse biomolecules and employed in biosensing to detect target analytes in biological samples. This proven concept was primarily used during the COVID-19 pandemic with gold-NP-based lateral flow assays (LFAs). Considering the gold price and its worldwide depletion, here we show that novel plasmonic NPs based on inexpensive metals, titanium nitride (TiN) and copper covered with a gold shell (Cu@Au), perform comparable to or even better than gold nanoparticles. After conjugation, these novel nanoparticles provided high figures of merit for LFA testing, such as high signals and specificity and robust naked-eye signal recognition. Since the main cost of Au NPs in commercial testing kits is the colloidal synthesis, our development with the Cu@Au and the laser-ablation-fabricated TiN NPs is exciting, offering potentially inexpensive plasmonic nanomaterials for various bioapplications. Moreover, our machine learning study showed that biodetection with TiN is more accurate than that with Au.

Tools for telehealth: A correlational analysis of app-based hearing testing.

Telehealth evaluation of hearing is rapidly evolving; however, the lack of consensus on the most accurate remote hearing test application has made hearing evaluation complicated. The objective of this study was to evaluate the correlation between the pure tone audiometry results obtained from app-based hearing testing programs and a traditional audiogram. A prospective within-subject and between-subject study design was used to correlate audiogram results between app-based hearing programs and a traditional audiogram. All participants completed a traditional audiogram, 1 commercial app-based test (ShoeBox), 2 consumer app-based tests (EarTrumpet and Hearing Test and Ear Age Test [HTEAT]), and a Hearing Handicap Inventory screening version (HHI-S). Testing was conducted in an acoustically controlled environment (traditional) and a quiet room (app-based hearing tests). A total of 39 participants were enrolled in the study (21 with normal hearing and 18 with hearing loss). In patients with normal hearing, only the commercial hearing testing app (ShoeBox) had a statistically significant pure tone average correlation in both ears with traditional audiometry (Right ear-r = 0.7, p = .005, Left ear-r = 0.66, p = .001). Both consumer and commercial apps had statistically significant correlations with both ears in patients with hearing loss (ranging from r = 0.62 to r = 0.9). Regarding accuracy within 10 dB of the pure tone average of the traditional audiogram of all tested ears, the commercial app-based test was accurate in 94% for all ears (normal and hearing loss), while consumer app-based tests were between 14% and 36% for all ears. The HHI-S indicated no hearing impairment in 95% of those with normal hearing and indicated hearing impairment in 89% of those with hearing loss. Commercial-grade app-based pure tone audiometry demonstrates overall strong correlation and accuracy with traditional audiometry. The HHI-S assessment remains a valid and useful tool to predict normal hearing and hearing impairment. 2.

Application of an improved wide–narrow-band hybrid ANC algorithm in a large commercial vehicle cabine

In recent years, active noise reduction technology has become a research hotspot. However, most active noise reduction technologies are developed based on passenger vehicles and are not implemented in large commercial vehicles. The large commercial vehicle sound field space is large, the traditional active noise reduction method is difficult to implement. In order to explore this problem, this paper proposes an improved active noise control method for commercial vehicles: (1) based on the traditional notch filter, a notch FxLMS algorithm based on speed smoothing is proposed; (2) on the basis of the traditional wire–narrowband hybrid ANC algorithm, the reference signal weighting technology is introduced into the wide-band subsystem, and the notch FxLMS algorithm based on speed smoothing is used as the narrowband subsystem, so as to propose an improved wire-narrowband hybrid ANC algorithm. With the help of MATLAB, the simulation model of the designed algorithm is established, and the collected commercial vehicle test noise data is used as the reference signal to simulate and verify the proposed algorithm. The results show that the proposed method has certain practicability.

Rapid and Sensitive Detection of Inactivated SARS-CoV-2 Virus via Fiber-Optic and Electrochemical Impedance Spectroscopy Based Aptasensors.

The development of rapid detection tools for viruses is vital for the prevention of pandemics and biothreats. Aptamers that target inactivated viruses are attractive for sensors due to their improved biosafety. Here, we evaluated a DNA aptamer (named as 6.9) that specifically binds to the inactivated SARS-CoV-2 virus with a low dissociation constant (KD = 9.6 nM) for the first time. Based on aptamer 6.9, we developed a fiber-optic evanescent wave (FOEW) biosensor. Inactivated SARS-CoV-2 and the Cy5.5-tagged short complementary strand competitively bound with the aptamer immobilized on the surface of the sensor. The detection of the inactivated SARS-CoV-2 virus was realized within six minutes with a limit of detection (LOD, S/N = 3) of 740 fg/mL. We also developed an electrochemical impedance aptasensor which exhibited an LOD of 5.1 fg/mL and high specificity. We further demonstrated that the LODs of the FOEW and electrochemical impedance aptasensors were, respectively, more than 1000 and 100,000 times lower than those of commercial colloidal gold test strips. We foresee that the facile aptamer isolation process and sensor design can be easily extended for the detection of other inactivated viruses.

Assessment of glial fibrillary acidic protein and anti-glial fibrillary acidic protein autoantibody concentrations and necrotising meningoencephalitis risk genotype in dogs with pug dog myelopathy.

Pugs commonly present with thoracolumbar myelopathy, also known as pug dog myelopathy (PDM), which is clinically characterised by progressive signs involving the pelvic limbs, no apparent signs of pain and, often, incontinence. In addition to meningeal fibrosis and focal spinal cord destruction, histopathology has confirmed lymphohistiocytic infiltrates in the central nervous system (CNS) in a considerable number of pugs with PDM. Lymphohistiocytic CNS inflammation also characterises necrotising meningoencephalitis (NME) in pugs. This study aimed to investigate the potential contribution of an immunological aetiology to the development of PDM. The concentrations of glial fibrillary acidic protein (GFAP) in serum and CSF and of anti-GFAP autoantibodies in CSF were measured with an ELISA. In addition, a commercial test was used for genetic characterisation of the dog leukocyte antigen class II haplotype, which is associated with NME susceptibility. This study included 87 dogs: 52 PDM pugs, 14 control pugs, four NME pugs and 17 dogs of breeds other than pugs that were investigated for neurological disease (neuro controls). Anti-GFAP autoantibodies were present in 15 of 19 (79%) of the PDM pugs tested versus six of 16 (38%) of the neuro controls tested (p = 0.018). All 18 PDM pugs evaluated had detectable CSF GFAP. Serum GFAP was detected in two of three (67%) of the NME pugs and in two of 11 (18%) of the control pugs but not in any of the 40 tested PDM pugs. Male pugs heterozygous for the NME risk haplotype had an earlier onset of clinical signs (70 months) compared to male pugs without the risk haplotype (78 months) (p = 0.036). The study was limited bythe lack of healthy dogs of breeds other than pugs and the small numbers of control pugs and pugs with NME. The high proportion of PDM pugs with anti-GFAP autoantibodies and high CSF GFAP concentrations provide support for a potential immunological contribution to the development of PDM.

Seroprevalence and co-infection of HBV, HCV, and HIV among patients visiting selected hospitals in Anyigba, Kogi State

ABSTRACT Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), and Human Immunodeficiency Virus (HIV) remain a public health challenge globally. This study determined the prevalence and coinfection of HBV, HCV, and HIV among patients visiting Maria Goretti Hospital, Grimard Catholic Hospital, and Good News Hospital Anyigba, Kogi State. In a cross-sectional study, sera samples collected from 400 consenting patients were screened for HBV, HCV, and HIV using commercial immunodiagnostic test kits. Of the 400 subjects, 12 (3.0%), 4 (1.0%), and 16 (4.0%) were infected with HBV, HCV, and HIV, respectively. One participant was co-infected with HCV and HIV, while none was simultaneously infected with HBV and HIV. Participants aged 11–20 years had higher hepatitis B-surface antigenemia, while ages 21–30 years and 31–40 years had higher prevalence of HCV and HIV, respectively. Contrary to HBV and HCV positivity, HIV seropositivity was significantly predicted by the ages of exposure (p = 0.002). Males and females were equally infected with HBV (3.0% each), while more males than females were infected with HCV (1.5%) and HIV (4.6%). However, the difference between the occurrence of viral infections and patients’ sex was not significant (p > 0.05). The single participants were more predisposed to HBV while the married subjects had more HCV and HIV mono-infection. However, neither the occurrence of HBV nor HCV or HIV was significantly predicted by the marital status of the individuals (p > 0.05). Subjects with no formal education had a higher positivity rate of HCV and HIV compared to other levels of education, while the tertiary level of education had higher exposure to HBsAg. Occupationally, students were more predisposed to HBV and HCV, while the unemployed participants were more predisposed to HIV. However, neither education nor the occupation of participants was significantly related to any of the viral infections (p > 0.05). Lack of knowledge of disease prevention significantly influenced the occurrence of HBV (p = 0.02), HCV (p = 0.04), and HIV (p = 0.04). Conclusively, the status of HBV, HCV, and HIV infection is low compared with findings of previous epidemiological studies in the area. However, the continuous circulation of the three viral infections and the high disease occurrence in the poorly informed participants suggest the need for increased public health education about infection control and prevention strategies in the area.

39 Using genetics and technology to improve swine robustness

Abstract Abstract Pig robustness has been an increasing concern in the global swine industry over the last 10 yr. This trend of increased mortality across all phases of production continues to worsen over time despite greater focus on research, management, and other production practices to control it. During this time, dramatic improvements in genetic progress have been realized through the selection of economically important traits. Historically, swine genetic companies have relied on data from their purebred animals reared in high-health and well-managed environments which has proven difficult to provide meaningful insight on variation in robustness traits. More recently, leading genetic companies have invested in large commercial test herds where crossbred pigs and breeding animals are individually identified with pedigree and genomic information. These pigs are reared under challenging and varying commercial conditions and mortality, removals, and reproductive efficiency data are gathered and used to generate crossbred breeding values. This type of data increases the accuracy of genetic estimates as well as creates new traits aimed at improving robustness and ease of management. Further investments have been made in precision livestock farming technologies such as computer vision which is quickly being developed due to the low cost of cameras and the ability to use edge computing to process images in real-time or to transfer images and data to the cloud. These developments are particularly compelling in the genetic improvement space when they can be paired with individual animal identification enabling novel traits to be measured in large populations more consistently and at a reduced cost compared with previous labor-intensive or cost-prohibitive methods. Early examples of this are the use of cameras to score feet and leg soundness in replacement gilts as lameness and associated feet and leg injuries are still one of the largest contributors to sow culls and mortality. This new digital feet and leg soundness scoring method has initially resulted in dramatically greater accuracy and heritability compared with conventional subjective human scoring methods. Other camera-based algorithms have been developed to track individual pigs and assess their daily behaviors and interactions. These developments are key to collecting new phenotypes for the selection of positive behavior traits for genetic selection compared with what was previously possible with human observer or scan sampling methods. Gene editing is another promising technology that has been used in human health and plant breeding but is now being widely researched in livestock. This technology has been used to create the first PRRS-resistant pigs to address one of the most costly diseases in the industry. New sources of data and technologies are key to creating and capturing existing and novel phenotypes on large populations of pigs in commercial environments to aid in the genetic improvement of non-conventional traits.

Concentrations of selected immunological parameters in the serum and processing fluid of suckling piglets and the serum and colostrum of their mothers

BackgroundBlood sampling from neonatal piglets is related to multiple disadvantages. Therefore, a new, alternative matrix is required to assess piglets’ early immune status efficiently. The present study aimed to assess the usefulness of processing fluid for determining selected piglets’ immune parameters. 264 pigs − 31 sows, 146 male piglets, and 87 female piglets from commercial indoor farrow-to-finish pig herd were included in this study. 264 serum, 31 colostrum, and 146 processing fluid samples were collected. Serum was collected from all animals, colostrum was collected from sows, and processing fluid was collected from male piglets only. Using commercial ELISA tests, the concentration of various immunoglobulins, cytokines, and acute phase proteins was assessed in each matrix. Statistical analyses were employed to determine differences in the concentration of measured indices between piglets’ serum and processing fluid and correlations in the concentration of tested indices between particular sets of matrices.ResultsStatistical analyses did not reveal significant differences in the IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ concentration between piglets’ serum and processing fluid (p > 0.05). A positive correlation (p < 0.05) regarding the concentration of some indices between processing fluid and samples collected from sows was also observed.ConclusionsProcessing fluid can be considered a promising alternative to blood for assessing some immunological indices in piglets, such as IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ, and, possibly, in the indirect assessment of some indices in lactating sows, including IgA, IL-1β, IL-4, IL-6, IL-8, IFN-γ, or Pig-MAP.

Viral SARS-CoV-2 rebound rates in linked commercial pharmacy-based testing and healthcare claims

Abstract Background Viral SARS-CoV-2 rebound (viral RNA rebound) is challenging to characterize in large cohorts due to the logistics of collecting frequent and regular diagnostic test results. Pharmacy-based testing data provides an opportunity to study the phenomenon in a large population, also enabling subgroup analyses. The current real-world evidence approach complements approaches focused on smaller, prospective study designs. Methods We linked RT-qPCR test data from national pharmacy-based testing to healthcare claims data via tokenization to calculate the cumulative incidence of viral RNA rebound within 28 days following positive test results in nirmatrelvir/ritonavir (NMV-r) treated and untreated individuals during the Omicron era (12/2021-11/2022) and prior to the Omicron era (10/2020-11/2-21). Findings Among 30,646 patients, the rate of viral RNA rebound was 3.5% (95% CI: 2.0-5.7%) in NMV-r treated infections compared to 1.5% (95% CI: 1.3-1.7%) in untreated infections during the Omicron era, and 1.9% (95% CI:1.7-2.1%) prior to the Omicron era. Viral RNA rebound in vaccinated (n=8,151), high-risk (n=4,411), or older patients (≥65yo, n=4,411) occurred at comparable rates to the overall cohort (range: 1.1-4.8%). Viral rebounds to high RNA levels in NMV-r treated infections occurred in 8% of viral rebounds, compared to 5-11% in untreated infections. Rates of hospitalization were comparable between patients with NMV-r treated infections with viral RNA rebound (0%) and untreated patients with viral RNA rebound (0-1.2%). Interpretation These findings related to viral RNA rebound are consistent with those of the EPIC-HR trial. Most occurrences of viral RNA rebound were associated with low viral RNA levels and viral RNA rebound progression to severe disease was not observed.

A low-cost design approach to vanadium redox flow test cell

Vanadium redox flow batteries (VRFB) will be essential in maximizing the large-scale use of renewable energy after achieving technical maturity. But as the test cells needed to conduct VRFB research are very expensive, the cost factor can prevent participation of developing countries in critical research. To address such issue, this work presents a novel low-cost design for laboratory VRFB cell yielding good performances. This design relies on sturdy ultra-high molecular weight polyethylene (UHMWPE) end plates with flow channels to provide the strength needed to seal other components while properly distributing electrolyte. This simpler design uses a reduced number of components compared to traditional cells, thus resulting in easier manufacturing and assembly while keeping costs low. This novel design approach achieved performances that are comparable to a commercial VRFB test cell in various testing conditions, but at roughly a third of the cost. Performances have been further improved by 3D printing end plates with optimized flow channels, with the added benefit of avoiding the complexity and costs of computer numerical control (CNC) milling. This novel design approach can thus be useful to many researchers in developing countries for their VRFB test.

Diagnostic accuracy of Afirma Gene Expression Classifier, Afirma Gene Sequencing Classifier, ThyroSeq v2 and ThyroSeq v3 for indeterminate (Bethesda III and IV) thyroid nodules: a meta-analysis.

The management of thyroid nodules with indeterminate cytology (ITN) is still a challenge. To evaluate the performance of commercial molecular tests for ITN, we performed this comprehensive meta-analysis. We performed an electronic search using PubMed/Medline, Embase and the Cochrane Library. Studies assessing the diagnostic accuracy of Afirma Gene Expression Classifier (GEC), Afirma Gene Sequencing Classifier (GSC), ThyroSeq v2 (TSv2) or ThyroSeq v3 (TSv3) in patients with ITN (only Bethesda category III or IV) were selected; Statistical analyses were performed by using Stata. 71 samples (GEC, n=38; GSC, n=16; TSv2, n=9; TSv3, n=8) in 53 studies, involving 6,490 fine needle aspirations (FNA) with ITN cytology with molecular diagnostics (GEC, GSC, TSv2 or TSv3) were included in the study. Meta-analysis showed the following pooled estimates: sensitivity 0.95 (95% CI, 0.94-0.97), specificity 0.35 (0.28-0.43), Positive Likelihood Ratio (LR+) 1.5 (1.3-1.6), and Negative Likelihood Ratio (LR-) 0.13 (0.09-0.19), with the best performance for TSv3 (area under the ROC curve 0.95 (0.93-0.96), followed by TSv2 (0.90 (0.87-0.92)), GSC (0.86 (0.82-0.88)) and GEC (0.82 (0.78-0.85)); with the best rule-out property for GSC (LR-, 0.07 (0.02-0.19)), followed by TSv3 (0.11 (0.05-0.24)) and GEC (0.16 (0.10-0.28); with the best rule-in for TSv2 (LR+, 2,9 (1.4-4.6)), followed by GSC (1.9 (1.6-2.4)). Meta-regression analysis revealed that study design, Bethesda category, and type of molecular test were independent factors. We showed that in patients with ITN, TSv3 has the best molecular diagnostic performance, followed by TSv2, GSC, and GEC. For rule-out malignancy, GSC, and for rule-in, TSV2 is superior to other tests.

Development and assessment of a multiepitope synthetic antigen for the diagnosis of Dengue virus infection

Immunodiagnostic tests for detecting dengue virus infections encounter challenges related to cross-reactivity with other related flaviviruses. Our research focuses on the development of a synthetic multiepitope antigen tailored for dengue immunodiagnostics. Selected dengue epitopes involved structural linearity and dissimilarity from the proteomes of Zika and Yellow fever viruses which served for computationally modeling the three-dimensional protein structure, resulting in the design of two proteins: rDME-C and rDME-BR. Both proteins consist of seven epitopes, separated by the GPGPG linker, and a carboxy-terminal 6 × -histidine tag. The molecular weights of the final proteins rDME-C and rDME-BR are 16.83 kDa and 16.80 kDa, respectively, both with an isoelectric point of 6.35. The distinguishing factor between the two proteins lies in the origin of their epitope sequences, where rDME-C is based on the reference dengue proteome, while rDME-BR utilizes sequences from prevalent Dengue genotypes in Brazil from 2008 to 2019. PyMol analysis revealed exposure of epitopes in the secondary structure. Successful expression of the antigens was achieved in soluble form and fluorescence experiments indicated a disordered structure. In subsequent testing, rDME-BR and rDME-C antigens were assessed using an indirect Elisa protocol against Dengue infected serum, previously examined with a commercial diagnostic test. Optimal concentrations for antigens were determined at 10 µg/mL for rDME-BR and 30 µg/mL for rDME-C, with serum dilutions ranging from 1:50 to 1:100. Both antigens effectively detected IgM and IgG antibodies in Dengue fever patients, with rDME-BR exhibiting higher sensitivity. Our in-house test showed a sensitivity of 77.3 % and 82.6 % and a specificity of 89.4 % and 71.4 % for rDME-C and rDEM-BR antigens. No cross-reactivity was observed with serum from Zika-infected mice but with COVID-19 serum samples. Our findings underscore the utility of synthetic biology in crafting Dengue-specific multiepitope proteins and hold promise for precise clinical diagnosis and monitoring responses to emerging Dengue vaccines.

Development and preliminary application of a quadruplex real-time PCR assay for differential detection of porcine circovirus 1-4 in Chengdu, China.

Porcine circovirus (PCV) typically causes severe immune suppression in pigs, leading to mixed clinical infections with various pathogens that can cause significant harm to the pig industry. PCV has four subgenotypes, with PCV4 being an emerging virus that requires investigation due to its potential for epidemic outbreaks. Therefore, there is a need to develop a method that can detect all four PCV strains simultaneously. In this study, four pairs of specific primers and TaqMan probes were designed based on the conserved sequence of the PCV1-4 ORF2 gene to establish a PCV1-4 TaqMan multiplex real-time quantitative PCR method. The novel method was compared to six commercial testing kits for its efficacy. Then, a total of 595 mixed samples of spleen and lymph node collected from 12 districts in Chengdu from July to December 2021 were tested using the novel method. The results showed that the novel PCV1-4 TaqMan multiplex real-time quantitative PCR detection method has satisfied specificity, sensitivity, and repeatability. The positive rates of PCV1, PCV2, and PCV3 in Chengdu were 2.18%, 31.60%, and 15.29%, respectively, while no positive PCV4 was detected. The mixed infection rate of PCV2 and PCV3 was 5.21%. Our novel method may be as a potential method for PCV1-4 detection. Currently, PCV2 is the main epidemic PCV subtype in Chengdu, while the potential threat of PCV4 should also be considered.

Clinical accuracy of instrument-based SARS-CoV-2 antigen diagnostic tests: a systematic review and meta-analysis

BackgroundDuring the COVID-19 pandemic, antigen diagnostic tests were frequently used for screening, triage, and diagnosis. Novel instrument-based antigen tests (iAg tests) hold the promise of outperforming their instrument-free, visually-read counterparts. Here, we provide a systematic review and meta-analysis of the SARS-CoV-2 iAg tests’ clinical accuracy.MethodsWe systematically searched MEDLINE (via PubMed), Web of Science, medRxiv, and bioRxiv for articles published before November 7th, 2022, evaluating the accuracy of iAg tests for SARS-CoV-2 detection. We performed a random effects meta-analysis to estimate sensitivity and specificity and used the QUADAS-2 tool to assess study quality and risk of bias. Sub-group analysis was conducted based on Ct value range, IFU-conformity, age, symptom presence and duration, and the variant of concern.ResultsWe screened the titles and abstracts of 20,431 articles and included 114 publications that fulfilled the inclusion criteria. Additionally, we incorporated three articles sourced from the FIND website, totaling 117 studies encompassing 95,181 individuals, which evaluated the clinical accuracy of 24 commercial COVID-19 iAg tests. The studies varied in risk of bias but showed high applicability. Of 24 iAg tests from 99 studies assessed in the meta-analysis, the pooled sensitivity and specificity compared to molecular testing of a paired NP swab sample were 76.7% (95% CI 73.5 to 79.7) and 98.4% (95% CI 98.0 to 98.7), respectively. Higher sensitivity was noted in individuals with high viral load (99.6% [95% CI 96.8 to 100] at Ct-level ≤ 20) and within the first week of symptom onset (84.6% [95% CI 78.2 to 89.3]), but did not differ between tests conducted as per manufacturer’s instructions and those conducted differently, or between point-of-care and lab-based testing.ConclusionOverall, iAg tests have a high pooled specificity but a moderate pooled sensitivity, according to our analysis. The pooled sensitivity increases with lower Ct-values (a proxy for viral load), or within the first week of symptom onset, enabling reliable identification of most COVID-19 cases and highlighting the importance of context in test selection. The study underscores the need for careful evaluation considering performance variations and operational features of iAg tests.

Recommendations for Improving the Use of Direct-To-Consumer Genetic Testing Databases in Law Enforcement Investigations

Prisoners are some of the most vulnerable populations in our society today, often relinquishing to the state everything from medical autonomy to family support. In a system where wrongful convictions are not unheard of, it is critical that we preserve the civil rights of investigative subjects wherever possible. Law enforcement agencies in the United States have increasingly begun using direct-to-consumer recreational genetic databases as a tool to enable forensic investigations. These commercial genetic testing companies analyze polymorphisms in DNA to isolate genealogical information, medical data, and other traits. Traditionally, law enforcement has been restricted to the use of matching repeating sequences in non-coding sections of DNA. However, through services like GEDMatch, law enforcement has gained access to single nucleotide polymorphism (SNP) markers from upwards of 25 million consumers throughout the world. This rapidly developing technology must be regulated to ensure that consumers, as well as blood relatives of consumers, are not unfairly targeted by investigations. With studies indicating that DNA is typically reported as the single most important piece of evidence to jurors, it is in the best interests of policymakers to ensure that the DNA data is both accurate and used fairly. First, we will provide a background of the technology and regulation of direct-to-consumer (DTC) genetic testing databases in law enforcement. Second, we propose a mechanism by which states might legislate the appropriate use of these databases by law enforcement by limiting access and enforcing a double-blind system. This paper lays out the background and policy proposal for a requirement of direct-to-consumer genetic testing companies to communicate the possibility and type of information shared with law enforcement, and a framework by which policymakers can ensure unbiased and minimally intrusive use by law enforcement.

HCV drug resistance mutations in HIV-infected patients

The aim of our work was to assess prevalence in the HCV drug resistance mutations in the NS3, NS5A, NS5B genes in HIV-infected patients. The material of the study was 157 blood plasma samples collected from HIV patients living in the Leningrad Region, with virological inefficiency of antiretroviral therapy. Samples were examined for the presence of anti-HCV antibodies and HCV RNA using commercial test systems in accordance with the manufacturer’s recommendations. In the case of detecting HCV RNA, amplification reaction was carried out using a set of primers co-flanking the NS3, NS5A, and NS5B genes. After sequencing the nucleotide sequences of the above genes, the virus subtype was determined. To assess HCV drug resistance mutations to direct-acting antiviral agents (DAAs), the Geno2pheno HCV resistance software (https:// hcv.geno2pheno.org) was used. The age of the patients varied from 18 to 65 and averaged 34.3±7.27 years. The number of males vs females prevailed comprising 71% and 28%, respectively. Antibodies against HCV were detected in 94.2% cases, whereas HCV RNA — in 98 (61.7%) HIV-infected persons. The distribution of HCV genotypes in this group was as follows: 1a — 7% (n = 7), 1b — 53% (n = 52), 2 — 2% (n = 2), 3a — 38% (n = 37). The results of determining the viral load varied from 5.3 × 103 to 2.3 × 108 IU/ml. The nucleotide sequence of all three regions NS3, NS5A, NS5B was determined in 73 samples. Analysis of drug resistance mutations in the NS3 region was performed for 82 HCV isolates of genotypes 1b (n = 46), 3a (n = 31), 1a (n = 4), 2 (n = 1). In total, the proportion of strains containing mutations in the NS3 region was 9.8%. The presence of significant amino acid substitutions in the NS5A region was tested for 83 HCV isolates of genotypes 1b (n = 47), 3a (n = 30), 1a (n = 6). The percentage of strains containing mutations in the NS5A region was 19.3%. Resistance mutations in the NS5B region were analysed for 87 HCV isolates of genotypes 1b (n = 48), 3a (n = 32), 2 (n = 2), 1a (n = 5). The proportion of strains containing mutations in the NS5B region was 11.5%. In the study group, mutations associated with hepatitis C virus resistance to DAAs in all regions were found in 16.6% (95% CI: 11.11–23.32%) cases (n = 26). In the NS3, NS5A and NS5B, there have been identified 6, 15 and 10 significant amino acid substitutions, respectively.

Association of polymorphic variants of cytokines genes, endothelial growth factor and matrix metalloproteinases with the development of uterine fibroids among russian women

One of the factors in the development of uterine fibroids is a genetic predisposition to its occurrence in some women, but the real molecular genetic mechanisms of this phenomenon remain unknown. Aim of the study was the distribution analysis of gene polymorphism of cytokines TNFα, IL-1β, IL-4, IL-6, IL-10, factors of angiogenesis (vascular endothelial growth factor, VEGF) and remodeling of extracellular matrix (metalloproteinases MMP2, MMP3, MMP9), which are associated with their levels. Material and methods. Genotyping was performed by real-time PCR using commercial test systems SYBR GreenI (Litech, Russia) and TaqMan (Syntol, Russia) in accordance with the instructions of the developer. Cytokine content in blood serum was determined by flow cytometry using microspheres coated with monoclonal antibodies to cytokines (Bio-Plex ProTM Human Cytokine 27-plex Assay), according to the instructions for Bio-Plex 200 (Bio-Rad Laboratories, USA).To evaluate the results obtained, in addition to the generally accepted methods of statistical processing for case – control studies, computational methods of bioinformatics were used for comparative analysis of the diagnostic value of various combined genetic traits. Results. It was shown that the maximum odds ratio value of uterine fibroids development are combined genetic traits that include representatives of all four regulatory factors: cytokines with pro-inflammatory activity, cytokines with anti-inflammatory activity, vascular endothelial growth factors and metalloproteinases (p = 0.002). Conclusions. The presented data reveal the real mechanisms of manifestation of the genetic predisposition of individual women to the uterine fibroids development, associated with the presence of polymorphism of certain genes in their genome, which provide features of the structure of cytokine networks with the predominance of certain activities in the regulation of tissue processes in the uterus. In addition to purely scientific interest, these results indicate the real possibility of their clinical application in the form of prognostic criteria with a certain level of prognostic significance.

Potential of a Bead-Based Multiplex Assay for SARS-CoV-2 Antibody Detection.

Serological assays for SARS-CoV-2 play a pivotal role in the definition of whether patients are infected, the understanding of viral epidemiology, the screening of convalescent sera for therapeutic and prophylactic purposes, and in obtaining a better understanding of the immune response towards the virus. The aim of this study was to investigate the performance of a bead-based multiplex assay. This assay allowed for the simultaneous testing of IgG antibodies against SARS-CoV-2 spike, S1, S2, RBD, and nucleocapsid moieties and S1 of seasonal coronaviruses hCoV-22E, hCoV-HKU1, hCoV-NL63, and hCoV-OC43, as well as MERS and SARS-CoV. We compared the bead-based multiplex assay with commercial ELISA tests. We tested the sera of 27 SARS-CoV-2 PCR-positive individuals who were previously tested with different ELISA assays. Additionally, we investigated the reproducibility of the results by means of multiple testing of the same sera. Finally, the results were correlated with neutralising assays. In summary, the concordance of the qualitative results ranged between 78% and 96% depending on the ELISA assay and the specific antigen. Repeated freezing-thawing cycles resulted in reduced mean fluorescence intensity, while the storage period had no influence in this respect. In our test cohort, we detected up to 36% of sera positive for the development of neutralising antibodies, which is in concordance with the bead-based multiplex and IgG ELISA.

The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants.

This study presents an evaluation of seventeen newly produced recombinant trivalent chimeric proteins (containing the same immunodominant fragment of SAG1 and SAG2 of Toxoplasma gondii antigens, and an additional immunodominant fragment of one of the parasite antigens, such as AMA1, GRA1, GRA2, GRA5, GRA6, GRA7, GRA9, LDH2, MAG1, MIC1, MIC3, P35, and ROP1) as a potential alternative to the whole-cell tachyzoite lysate (TLA) used in the detection of infection in small ruminants. These recombinant proteins, obtained by genetic engineering and molecular biology methods, were tested for their reactivity with specific anti-Toxoplasma IgG antibodies contained in serum samples of small ruminants (192 samples of sheep serum and 95 samples of goat serum) using an enzyme-linked immunosorbent assay (ELISA). The reactivity of six recombinant trivalent chimeric proteins (SAG1-SAG2-GRA5, SAG1-SAG2-GRA9, SAG1-SAG2-MIC1, SAG1-SAG2-MIC3, SAG1-SAG2-P35, and SAG1-SAG2-ROP1) with IgG antibodies generated during T. gondii invasion was comparable to the sensitivity of TLA-based IgG ELISA (100%). The obtained results show a strong correlation with the results obtained for TLA. This suggests that these protein preparations may be a potential alternative to TLA used in commercial tests and could be used to develop a cheaper test for the detection of parasite infection in small ruminants.