Development and clinical evaluation of a rapid antibody lateral flow assay for the diagnosis of SARS-CoV-2 infection.

BACKGROUND: The novel coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which has quickly spread worldwide since its outbreak in December 2019. One of the primary measures for controlling the spread of SARS-CoV-2 infection is an accurate assay for its diagnosis. SARS-CoV-2 real-time PCR kits suffer from some limitations, including false-negative results in the clinic. Therefore, there is an urgent need for the development of a rapid antibody test kit for COVID-19 diagnosis. METHODS: The nuclear capsid protein (N) and spike protein 1 (S1) fragments of SARS-CoV-2 were expressed in Escherichia coli, and rapid antibody-based tests for the diagnosis of SARS-CoV-2 infection were developed. To evaluate their clinical applications, the serum from COVID-19 patients, suspected COVID-19 patients, recovering COVID-19 patients, patients with general fever or pulmonary infection, doctors and nurses who worked at the fever clinic, and health professionals was analyzed by the rapid antibody test kits. The serum from patients infected with Mycoplasma pneumoniae and patients with respiratory tract infection was further analyzed to test its cross-reactivity with other respiratory pathogens. RESULTS: A 47 kDa N protein and 67 kDa S1 fragment of SARS-CoV-2 were successfully expressed, purified, and renatured. The rapid antibody test with recombinant N protein showed higher positive rate than the rapid IgM antibody test with recombinant S1 protein. Clinical evaluation showed that the rapid antibody test kit with recombinant N protein had 88.56 % analytical sensitivity and 97.42 % specificity for COVID-19 patients, 53.48 % positive rate for suspected COVID-19 patients, 57.14 % positive rate for recovering COVID-19 patients, and 0.5-0.8 % cross-reactivity with other respiratory pathogens. The analytical sensitivity of the kit did not significantly differ in COVID-19 patients with different disease courses (p < 0.01). CONCLUSIONS: The rapid antibody test kit with recombinant N protein has high specificity and analytical sensitivity, and can be used for the diagnosis of SARS-CoV-2 infection combined with RT-PCR.

A prospective study on associations between superoxide dismutase gene polymorphisms and antituberculosis drug-induced liver injury in a Chinese Han population.

BACKGROUND: Antituberculosis drug-induced liver injury (ATDILI) is increasing globally and, hence, it is crucial to predict its risk in the clinical management of antituberculosis therapy. As a major antioxidant, superoxide dismutase (SOD) is mainly responsible for providing defence against oxidative stress, which is involved in ATDILI. The present study aimed to investigate the associations between polymorphisms in SOD genes, including Cu/ZnSOD (SOD1), mitochondrial manganese SOD (MnSOD or SOD2) and extracellular SOD (SOD3), as well as the susceptibility to ATDILI in a Chinese Han population. METHODS: In total, 1060 Chinese Han subjects highly suspected to have tuberculosis (TB) were prospectively enrolled from West China Hospital of Sichuan University. Overall, 746 subjects comprising 118 ATDILI and 628 ATD-tolerant TB patients were eligible and were genotyped for seven single-nucleotide polymorphisms in three SOD genes (SOD1: rs4816407 and rs1041740; SOD2: rs4880; SOD3: rs699473, rs2536512, rs2855262 and rs8192290). RESULTS: Logistic regression analysis showed that none of the seven genetic variants in the three SOD genes were significantly associated with susceptibility to ATDILI in the Chinese Han population after Bonferroni correction, except for a potential association for the SOD2 rs4880 A>G (G allele, p = 0.190, odds ratio = 1.53, 95% confidence interval = 1.05-2.23; GG genotype, p = 0.155). CONCLUSIONS: The promising application of single-nucleotide polymorphisms in the SOD1, SOD2 and SOD3 genes as genetic markers for ATDILI is challenged, and further studies are needed with larger sample sizes and different ethnicities, especially for SOD2 rs4880.

Comparison of the community-acquired pneumonia and COVID-19 at the early stage: findings from two cohort studies.

BACKGROUND: The current focus is largely on whole course medical management of coronavirus disease-19 (COVID-19) with real-time polymerase chain reaction (RT-PCR) and radiological features, while the mild cases are usually missed. Thus, combination of multiple diagnostic methods is urgent to understand COVID-19 fully and to monitor the progression of COVID-19. METHODS: laboratory variables of 40 mild COVID-19 patients, 30 patients with community-acquired pneumonia (CAP) and 32 healthy individuals were analyzed by principal component analysis (PCA), Kruskal test, Procrustes test, the vegan package in R, CCA package and receiver operating characteristic to investigate the characteristics of the laboratory variables and their relationships in COVID-19. RESULTS: The correlations between the laboratory variables presented a variety of intricate linkages in the COVID-19 group compared with the healthy group and CAP patient group. The prediction probability of the combination of lymphocyte count (LY), eosinophil (EO) and platelets (PLT) was 0.847, 0.854 for the combination of lactate (LDH), creatine kinase isoenzyme (CK-MB), and C-reactive protein (CRP), 0.740 for the combination of EO, white blood cell count (WBC) and neutrophil count (NEUT) and 0.872 for the combination of CK-MB and P. CONCLUSIONS: The correlations between the laboratory variables in the COVID-19 group could be a unique characteristic showing promise as a method for COVID-19 prediction and monitoring progression of COVID-19 infection.

MicroRNA-27a restrains the immune response to mycobacterium tuberculosis infection by targeting IRAK4, a promoter of the NF-κB pathway.

This study aimed to explore the influence of microRNA-27a on immune response to mycobacterium tuberculosis (Mtb) and the molecular mechanism. MiRNA-27a was screened one of the downregulated miRNAs in Mtb infected macrophages. The concentrations of IFN-γ, IL-ß, IL-6, and TNF-α in THP-1 macrophages after infection with Mtb and simultaneous transfection with miR-27a mimics or inhibitor were determined by ELISA. Colony-forming unit (CFU) assay was used to determine the survival situation of THP-1 infected with Mtb after transfection with miR-27a mimics or inhibitor. We used luciferase reporter assay and western blotting to study the relationship between miR-27a and IRAK4. MiR-27a was found differential expressed in Mtb infected macrophages, and the expressions of miR-27a in Mtb-infected THP-1 were remarkably downregulated with the increase of time and dose. Compared with the control, the levels of IFN-γ, IL-ß, IL-6, and TNF-α were enhanced after macrophages infected with Mtb, while further transfection of miR-27a mimics abolished the increase. IRAK4 was found the target gene of miR-27a and transfection of miR-27a mimics decreased the relative level of IRAK4. The concentration of IFN-γ, IL-ß, IL-6, and TNF-α in Mtb infected macrophages were reduced significantly after transfection of miR-27a mimics, while simultaneous transfection of pcDNA-IRAK4 abolished the decrease, which is the upstream molecular of NF-κB. In conclusion, miR-27a plays a key role in immune response to Mtb infection and intervening on miR-27a may be an effective way to treat TB.

First insight into the genotypic diversity of clinical Mycobacterium tuberculosis isolates from Gansu Province, China.

BACKGROUND: Investigations of Mycobacterium tuberculosis genetic diversity in China have indicated a significant regional distribution. The aim of this study was to characterize the genotypes of clinical M. tuberculosis isolates obtained from Gansu, which has a special geographic location in China. METHODOLOGY/PRINCIPAL FINDINGS: A total of 467 clinical M. tuberculosis strains isolated in Gansu Province were genotyped by 15-locus mycobacterial interspersed repetitive units-variable number tandem repeats (MIRU-VNTR) and spoligotyping. The results showed that 445 isolates belonged to six known spoligotype lineages, whereas 22 isolates were unknown. The Beijing genotype was the most prevalent (87.58%, n = 409), while the shared type 1 was the dominant genotype (80.94%, n = 378). The second most common lineage was the T lineage, with 25 isolates (5.35%), followed by the H lineage with 5 isolates (1.07%), the MANU family (0.64%, 3 isolates), the U family (0.43%, 2 isolates) and the CAS lineage with 1 isolate (0.21%). By using the VNTR15China method, we observed 15 groups and 228 genotypes among the 467 isolates. We found no association between the five larger groups (including the Beijing genotype) and sex, age, or treatment status, and there was no noticeable difference in the group analysis in different areas. In the present study, seven of the 15 MIRU-VNTR loci were highly or moderately discriminative according to their Hunter-Gaston discriminatory index. CONCLUSIONS/SIGNIFICANCE: The Beijing genotype is the predominant genotype in Gansu province. We confirm that VNTR15China is suitable for typing Beijing strains in China and that it has a better discriminatory power than spoligotyping. Therefore, the use of both methods is the most suitable for genotyping analysis of M. tuberculosis.