Multiplex real-time RT-PCR method for the diagnosis of SARS-CoV-2 by targeting viral N, RdRP and human RP genes.

Corona Virus Disease 2019 (COVID-19) is a disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This pandemic has brought the world to a standstill and threatened human lives. Many methods are known to date to detect this virus. Due to their relative sensitivity, polymerase chain reaction (PCR)-based assays are the most frequently applied and considered the gold standard. However, due to the rapid mutation rate of the viral genome and the emergence of new variants, existing protocols need to be updated and improved. Designing a fast and accurate PCR-based assay is of great importance for the early detection of this virus and more efficient control of the spread of this disease. This study describes a fast, reliable, easy-to-use, and high-throughput multiplex SARS-CoV-2 RT-PCR detection method. The assay was designed to detect two viral genes (N and RdRP) and a human gene (RP) simultaneously. The performance and the sensitivity of the assay were tested in 28 SARS-CoV-2 positive samples and compared with commercial kits, which showed 100% positive percent agreement with a limit of detection (LOD) value of 1.40 and 0.81 copies/µL or 35.13 and 20.31 copies/reaction for RdRP and N genes, respectively. The current assay is found accurate, reliable, simple, sensitive, and specific. It can be used as an optimized SARS-CoV-2 diagnostic assay in hospitals, medical centers, and diagnostic laboratories as well as for research purposes.

Designing of Co0.5Ni0.5GaxFe2-xO4 (0.0 ≤ x ≤ 1.0) Microspheres via Hydrothermal Approach and Their Selective Inhibition on the Growth of Cancerous and Fungal Cells.

The current study offers an efficient design of novel nanoparticle microspheres (MCs) using a hydrothermal approach. The Co0.5Ni0.5GaxFe2-xO4 (0.0 ≤ x ≤ 1.0) MCs were prepared by engineering the elements, such as cobalt (Co), nickel (Ni), iron (Fe), and gallium (Ga). There was a significant variation in MCs' physical structure and surface morphology, which was evaluated using energy dispersive X-ray analysis (EDX), X-ray diffractometer (XRD), high-resolution transmission electron microscopy (HR-TEM), and scanning electron microscope (SEM). The anti-proliferative activity of MCs was examined by MTT assay and DAPI staining using human colorectal carcinoma cells (HCT-116), human cervical cancer cells (HeLa), and a non-cancerous cell line-human embryonic kidney cells (HEK-293). Post 72 h treatment, MCs caused a dose dependent inhibition of growth and proliferation of HCT-116 and HeLa cells. Conversely, no cytotoxic effect was observed on HEK-293 cells. The anti-fungal action was assessed by the colony forming units (CFU) technique and SEM, resulting in the survival rate of Candida albicans as 20%, with severe morphogenesis, on treatment with MCs x = 1.0. These findings suggest that newly engineered microspheres have the potential for pharmaceutical importance, in terms of infectious diseases and anti-cancer therapy.

Anti-microbial and anti-cancer activities of Mn0.5Zn0.5DyxFe2-xO4 (x ≤ 0.1) nanoparticles.

Combining two or more nanoparticles is a promising approach. Previously we have reported synthesis of nanoparticles Dysprosium (Dy) substituted with manganese (Mn) zinc (Zn) by using ultrasonication method. The five different nanoparticles (NPs) Mn0.5Zn0.5DyxFe2-xO4 (x ≤ 0.1) have been structurally and morphologically characterized but there is no report on the biological application of these NPs. In the present study, we have examined the anti-cancer, anti-bacterial, and anti-fungal activities of Mn0.5Zn0.5DyxFe2-xO4 (x ≤ 0.1) NPs. Human colorectal carcinoma cells (HCT-116) were tested with different concentrations of NPs by using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. In addition, the impact of NPs was also examined on normal cells such as human embryonic kidney cells, HEK-293. After 48 h of treatment, Mn0.5Zn0.5DyxFe2-xO4 NPs (x = 0.02, 0.04 and 0.06) showed no inhibitory action on cancer cell's growth and proliferation, whereas Mn0.5Zn0.5DyxFe2-xO4 NPs (x = 0.08 and 0.1) showed profound inhibitory action on cancer cell's growth and proliferation. However, the treatment of Mn0.5Zn0.5DyxFe2-xO4 NPs on the normal cells (HEK-293) did not show cytotoxic or inhibitory action on HEK-293 cells. The treatment of Mn0.5Zn0.5DyxFe2-xO4 NPs (x ≤ 0.1) also inhibited both the bacteria (Escherichia coli ATCC35218 and Staphylococcus aureus) with lowest MIC and MBC values of 4 and 8 mg/mL and fungus (Candida albicans) with MIC and MFC values of 4 and 8 mg/mL on treatment with x = 0.08 and 0. 1.

Synthesis, Characterization, Anti-Cancer Analysis of Sr0.5Ba0.5DyxSmxFe8-2xO19 (0.00 ≤ x ≤ 1.0) Microsphere Nanocomposites.

There is enormous interest in combining two or more nanoparticles for various biomedical applications, especially in anti-cancer agent delivery. In this study, the microsphere nanoparticles were prepared (MSNPs) and their impact on cancer cells was examined. The MSNPs were prepared by using the hydrothermal method where strontium (Sr), barium (Ba), dysprosium (Dy), samarium (Sm), and iron oxide (Fe8-2xO19) were combined, and dysprosium (Dy) and samarium (Sm) was substituted with strontium (Sr) and barium (Ba), preparing Sr0.5Ba0.5DyxSmxFe8-2xO19 (0.00 ≤ x ≤ 1.0) MSNPs. The microspheres were characterized by X-ray powder diffraction (XRD), high-resolution transmission electron microscopy (HR-TEM), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX) techniques. The diffraction pattern of nanohexaferrites (NHFs) reflected the signature peaks of the hexagonal structure. The XRD revealed a pure hexagonal structure without any undesired phase, which indicated the homogeneity of the products. The crystal size of the nanoparticles were in the range of 22 to 36 nm by Scherrer's equation. The SEM of MSNPs showed a semi-spherical shape with a high degree of aggregation. TEM and HR-TEM images of MSNPs verified the spherical shape morphology and structure that approved an M-type hexaferrite formation. The anti-cancer activity was examined on HCT-116 (human colorectal carcinoma) and HeLa (cervical cancer cells) using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and post-48 h treatment of MSNPs caused a dose-dependent inhibition of HCT-116 and HeLa cell proliferation and growth. Conversely, no significant cytotoxic effect was observed on HEK-293 cells. The treatments of MSNPs also induced cancer cells DNA disintegration, as revealed by 4',6-diamidino-2-phenylindole (DAPI) staining. Finally, these findings suggest that synthesized MSNPs possess potential inhibitory actions on cancerous cells without harming normal cells.