ABSTRACT
INTRODUCTION: COVID-19 is a worldwide public health threat. Diagnosis by RT-PCR has been employed as the standard method to confirm viral infection. Sample pooling testing can optimize the resources by reducing the workload and reagents shortage, and be useful in laboratories and countries with limited resources. This study aims to evaluate SARS-CoV-2 detection by sample pooling testing in comparison with individual sample testing. MATERIALS AND METHODS: We created 210 pools out of 245 samples, varying from 4 to 10 samples per pool, each containing a positive sample. We conducted detection of SARS-CoV-2-specific RdRp/E target sites. RESULTS: Pooling of three samples for SARS-CoV-2 detection might be an efficient strategy to perform without losing RT-PCR sensitivity. CONCLUSIONS: Considering the positivity rate in Dominican Republic and that larger sample pools have higher probabilities of obtaining false negative results, the optimal sample size to perform a pooling strategy shall be three samples.
ABSTRACT
Nucleic acid detection has been one of the most valued tools in point-of-care diagnostics from life science, agriculture, food safety and environmental surveillance, because of its high sensitivity, great specificity and simple operation. Since polymerase chain reactions (PCR) were discovered, more and more researchers attach importance to exploring ultrafast nucleic acid amplification methods for further expediting the process of detection and curbing infectious diseases' high spread rate, especially after the coronavirus disease 2019 (COVID-19) worldwide pandemic event. Nowadays, nanotechnology as one of the most cut-ting-edge technologies has aroused growing attention. In this review, we describe new advances in na-notechnology research for ultrafast nucleic acid amplification. We have introduced commonly used nanotechnologies, namely nanofluidics, nanoporous materials, nanoparticles and so on. Recent advances in these nanotechnologies for ultrafast sample pretreatments, accelerated enzymatic amplification and rapid heating/cooling processes was summarized. Finally, challenges and perspectives for the future applications of ultrafast nucleic acid amplification are presented.(c) 2022 Elsevier Ltd. All rights reserved.
ABSTRACT
The chicken is rich in various proteins, fatty acids, polysaccharides, trace elements, and other human essential nutrients that contribute to its high nutritional value. In this study, the expression levels of nutrition-related genes (acetyl-CoA acyltransferase, ACAA) of native chicken breeds were investigated. The level of GgalACAA1-2 transcripts expression in the liver of chicken was significantly higher than that of muscle and heart. Moreover, three protein extracts were isolated from the muscle, heart, and liver tissues from the chicken, and their nutritional function was evaluated in the present study. These protein extracts had excellent DPPH and hydroxyl radical scavenging capacities and exhibited significant superoxide anion scavenging ability. Moreover, the protein extracts of muscle tissue showed an important mouse splenocyte proliferation activity and could be used as an immunomodulator of natural origin. In addition, this report presented an automatic visual inspection of chicken viscera using the active contour algorithms and the image processing method for eviscerating by the parallel robot. The recognition and positioning rate of chicken viscera obtained by the proposed method could reach 96.45%. These methods provided basic data for automated poultry slaughter and segmentation, avoiding unnecessary health risks by a pathogenic microorganism, such as avian influenza, Newcastle disease virus, and coronavirus. Moreover, the internal organs of the chicken could be fully harvested by the image segmentation of automatic evisceration, which also facilitated the processing value of these internal organs as by-products of poultry.
ABSTRACT
The COVID-19 pandemic, caused by severe acute respiratory syndrome corona virus 2 (SARS-CoV-2), has become a severe global health problem affecting almost every country in the world. Compared to other coronaviruses, SARS-CoV-2 is considered to be more infectious thereby leading to a rapid spread of this disease across the world. The effective control of this disease relies on timely diagnosis, proper isolation, contact tracing of the infected people and segregation of vulnerable group from potential contamination. Currently, the gold standard diagnostic test for COVID-19 is real-time reverse transcriptase polymerase chain reaction (RT-PCR) using nasopharyngeal swab (NPS). However, NPS collection has several shortcomings. Besides requiring an active involvement of healthcare personnel and personal protective equipment (PPE), NPS collection is uncomfortable for the patient as it can induce coughing, gagging, vomiting and even bleeding. Evidence from current studies indicates that saliva has a potential to be useful as an alternative biological sample for COVID-19 diagnosis. Indeed, saliva as a biological sample offers several advantages over NPS. Saliva collection is better accepted by patients, it can be self-collected and does not require PPE and active involvement of healthcare personnel. Moreover, preliminary results indicate that the sensitivity and specificity of saliva for COVID-19 diagnosis is similar to that of NPS. This summarizes recent observations in the field and discusses the potential use of saliva for COVID-19 diagnosis. Copyright © 2020, Kathmandu University. All rights reserved.
ABSTRACT
BACKGROUND: The progression of infectious diseases depends on the characteristics of a patient's innate immunity, and the efficiency of an immune system depends on the patient's genetic factors, including SNPs in the TLR genes. In this pilot study, we determined the frequency of alleles in these SNPs in a subset of patients with pneumonia. METHODS: This study assessed six SNPs from TLR genes: rs5743551 (TLR1), rs5743708, rs3804100 (TLR2), rs4986790 (TLR4), rs5743810 (TLR6), and rs3764880 (TLR8). Three groups of patients participated in this study: patients with pneumonia in 2019 (76 samples), patients with pneumonia caused by SARS-CoV-2 in 2021 (85 samples), and the control group (99 samples). RESULTS: The allele and genotype frequencies obtained for each group were examined using four genetic models. Significant results were obtained when comparing the samples obtained from individuals with pneumonia before the spread of SARS-CoV-2 and from the controls for rs5743551 (TLR1) and rs3764880 (TLR8). Additionally, the comparison of COVID-19-related pneumonia cases and the control group revealed a significant result for rs3804100-G (TLR2). CONCLUSIONS: Determining SNP allele frequencies and searching for their associations with the course of pneumonia are important for personalized patient management. However, our results need to be comprehensively assessed in consideration of other clinical parameters.
ABSTRACT
Infectious bronchitis (IB) is a highly contagious viral disease of chickens caused by IB virus (IBV) that can cause substantial economic losses in the poultry industry. IBV variant infections have been continuously reported since the initial description in the 1930s. QX-like IBVs are the predominant circulating genotype globally. A homologous QX vaccine has superior protection efficacy compared with that of other available vaccines, and the combination of Massachusetts (Mass)-like and QX-like strains is being used to combat QX-like IBV infections. Inoculation of embryonated chicken eggs is the standard method for the titration of IBV, and the titer is expressed as 50% egg infectious dose (EID50). However, this method cannot effectively distinguish or quantify different genotypic strains in a mixture of different viruses, especially in the absence of neutralizing monoclonal antibodies. In this study, quantitative real-time PCR (RT-qPCR) was applied using specific primers for the QX- and Mass-like strains to quantitate IBV infection and for comparison with the conventional virus titration quantitative method. A strong positive correlation was observed between RT-qPCR cycle threshold values and the different EID50 concentrations. This method was further used to titrate bivalent IB vaccines, and the amount of individual genotype virus was determined based on specific primers. Thus, this RT-qPCR assay may be used as a highly specific, sensitive, and rapid alternative to the EID50 assay for titering IBVs.