RESUMO
To rapidly identify individuals infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and control the spread of coronavirus disease (COVID-19), there is an urgent need for highly sensitive on-site virus detection methods. A clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas)-based molecular diagnostic method was developed for this purpose. Here, a CRISPR system-mediated lateral flow assay (LFA) for SARS-CoV-2 was established based on multienzyme isothermal rapid amplification, CRISPR-Cas13a nuclease, and LFA. To improve the limit of detection (LoD), the crispr RNA, amplification primer, and probe were screened, in addition to concentrations of various components in the reaction system. The LoD of CRISPR detection was improved to 0.25 copy/µl in both fluorescence- and immunochromatography-based assays. To enhance the quality control of the CRISPR-based LFA method, glyceraldehyde-3-phosphate dehydrogenase was detected as a reference using a triple-line strip design in a lateral flow strip. In total, 52 COVID-19-positive and 101 COVID-19-negative clinical samples examined by reverse transcription polymerase chain reaction (RT-PCR) were tested using the CRISPR immunochromatographic detection technique. Results revealed 100% consistency, indicating the comparable effectiveness of our method to that of RT-PCR. In conclusion, this approach significantly improves the sensitivity and reliability of CRISPR-mediated LFA and provides a crucial tool for on-site detection of SARS-CoV-2.
Assuntos
COVID-19 , Proteínas Associadas a CRISPR , COVID-19/diagnóstico , Proteínas Associadas a CRISPR/genética , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA , Reprodutibilidade dos Testes , SARS-CoV-2/genética , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: We aimed to systematically evaluate the effectiveness of the currently available mRNA vaccines and boosters for the Omicron variant. METHODS: We searched for literature published on PubMed, Embase, Web of Science and preprint servers (medRxiv and bioRxiv) from January 1, 2020 to June 20, 2022. The pooled effect estimate was calculated by the random-effects model. RESULTS: We selected 34 eligible studies in the meta-analysis from 4336 records. For the 2-dose vaccinated group, the mRNA vaccine effectiveness (VE) was 34.74%, 36%, and 63.80% against any Omicron infection, symptomatic infection and severe infection, respectively. For the 3-dose vaccinated group, the mRNA VE was 59.80%, 57.47%, and 87.22% against any infection, symptomatic infection and severe infection. For the 3-dose vaccinated group, the relative mRNA VE was 34.74%, 37.36%, and 63.80% against any infection, symptomatic infection and severe infection. Six months after the 2-dose vaccination, VE with any infection, symptomatic infection, and severe infection decreased to 33.4%, 16.79%, and 60.43%. Three months after the 3-dose vaccination, VE for any infection and severe infection decreased to 55.39% and 73.39%. CONCLUSIONS: Two-dose mRNA vaccines failed to provide sufficient protection against any Omicron infection and symptomatic infection, while 3-dose mRNA vaccines continued to provide effective protection after 3 months.
Assuntos
Vacinação , Vacinas de mRNA , Humanos , Imunização Secundária , RNA Mensageiro/genéticaRESUMO
Although numerous COVID-19 vaccines are effective against COVID-19 infection and variants of concern (VOC) in the real world, it is imperative to obtain evidence of the corresponding vaccine effectiveness (VE). This study estimates the real-world effectiveness of the BNT162b2 and mRNA-1273 vaccines against COVID-19 infection and determines the influence of different virus variants on VE by using test-negative design (TND) studies. We systematically searched for published articles on the efficacy of BNT162b2 and mRNA-1273 against COVID-19 infection. Two researchers independently selected and extracted data from eligible studies. We calculated the VE associated with different vaccine types, SARS-CoV-2 variants, and vaccination statuses, using an inverse variance random-effects model. We selected 19 eligible studies in the meta-analysis from 1651 records. For the partially vaccinated group, the VE of BNT162b2 and mRNA-1273 was 61% and 78% against COVID-19 infection, respectively. For the completely vaccinated group, the VE of BNT162b2 and mRNA-1273 was 90% and 92% against COVID-19 infection, respectively. During subgroup analyses, the overall VE of BNT162b2 and mRNA-1273 against the Delta variant was 53% and 71%, respectively, for the partially vaccinated group; the respective VE values were 85% and 91% for the fully vaccinated group. Irrespective of the BNT162b2 or mRNA-1273 vaccines, the Delta variant significantly weakened vaccine protection for the partially vaccinated group, while full vaccination was highly effective against COVID-19 infection and various VOC. The mRNA-1273 vaccine is more effective against COVID-19 infection and VOC than the BNT162b2 vaccine, especially for the partially vaccinated group. Overall, the results provide recommendations for national and regional vaccine policies.
RESUMO
BACKGROUND: Food allergy has been a significant public health issue with growing severity, prevalence and limited treatments. The neutrophil-activating protein A subunit (NapA) of Helicobacter pylori has been shown to have therapeutic potential in allergic diseases. METHODS: The NapA expression efficiency of recombinant Lactococcus lactis(L.lactis) were determined. The effects of recombinant bacterium on food allergy in Balb/c mice were also investigated. RESULTS: NapA were delivered and expressed efficiently via L. lactis. The engineered bacterium ameliorated food allergy symptoms (acute diarrhea and intestinal inflammation) and decreased serum histamine levels. In addition, the secretion of OVA-specific IgG2a, IFN-γ was promoted and the level of IL-4, OVA-specific IgE was restrained. CONCLUSIONS: The recombinant strain may attenuate food allergy in mice through immune regulatory effect, which may be a promising approach for preventing or treating food allergy.