The outbreak of the new human coronavirus SARS-CoV-2 (also known as 2019-nCoV) continues to increase globally. The real-time reverse transcription polymerase chain reaction (rRT-PCR) is the most used technique in virus detection. However, possible false-negative and false-positive results produce misleading consequences, making it necessary to improve existing methods. Here, we developed a multiplex rRT-PCR diagnostic method, which targets two viral genes (RdRP and E) and one human gene (RP) simultaneously. The reaction was tested by using pseudoviral RNA and human target mRNA sequences as a template. Also, the protocol was validated by using 14 clinical SARS-CoV-2 positive samples. The results are in good agreement with the CDC authorized Cepheid`s Xpert® Xpress SARS-CoV-2 diagnostic system (100%). Unlike single gene targeting strategies, the current method provides the amplification of two viral regions in the same PCR reaction. Therefore, an accurate SARS-CoV-2 diagnostic assay was provided, which allows testing of 91 samples in 96-well plates in per run. Thanks to this strategy, fast, reliable, and easy-to-use rRT-PCR method is obtained to diagnose SARS-CoV-2.
Biocompatible SnO 2 NPs, were synthesized by simple and facile ultrasonic technique, and further studied for antibacterial, anticandidal and invitro cytotoxic activity. The obtained size of synthesized SnO 2 NPs, was 5-30 nm and characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM). The evaluation of antibacterial and anticandidal activity of the SnO 2 NPs, was done by determining the minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC/MFC), against Escherichia coli and Candida albicans, respectively. The MIC of 0.5 mg/ml and MBC of > 1 mg/ml for E. coli, while as the MIC of 8 mg/ml and MFC > 16 mg/ml for C. albicans, was obtained. Maximum activity for both, bacteria and Candida was achieved, however, to the best of our knowledge, this is the first report of anticandidal activity exhibited by SnO 2 NPs. Treatment of synthesized SnO 2 NPs against the colorectal (HCT-116) cancer cell line, decreased the cell survivability in a dose-dependent manner, as lower dose showed decrease of 73.00%, whereas, higher dosage caused a significant 31.34% decrease in the cell viability. The obtained results confirmed, that the prepared SnO 2 NPs can be the future broad-spectrum antibacterial, anticandidal and anticancer agent, and can be further explored for application in the biomedical field.promising potential for use in the pharmaceutical and biomedical applications. Supporting information summaryThe experimental section includes the detailed procedure for synthesis, characterization, antibacterial, anticandidal and cytotoxic activities of Tin Oxide NPs (SnO 2 NPs), which can be find in supporting information.
The establishment of a benign system for the nanoparticle (NPs) synthesis, is a key in nanotechnology for the environmental and health care industries. Therefore, enrichment of novel biological systems for the green synthesis is in significant demand, to lift up these compounds in the biomedical industries. The present work, reports the green synthesis of ZnO NPs, employing a novel thermophile, identified as Bacillus haynesii (GeneBank: MG822851) isolated from the leaf of date palm plant (Phoenix dactylifera), as an eco-friendly nanobiofactory. Physiochemical characterization of ZnO NPs (50 ± 5 nm in size), was achieved by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), diffuse reflectance UV-Visible spectroscopy (DR UV-Vis spectroscopy), Thermogravimetry analysis (TGA), scanning electron microscopy (SEM) and transmissiom electron microscopy (TEM). The morphogenesis and antimicrobial activity of synthesized ZnO NPs, was studied by evaluating the minimum inhibitory/bactericidal concentration (MIC&MBC) against Escherchia coli (8 and 16 mg/mL) and Staphylococcus aureus (4 and 8 mg/mL), respectively. The present study encourages the use of B. haynesii for the green synthesis of ZnO NP. To the best of our knowledge, this is the first report on the study of thermophilic, B. haynesii for green synthesis of NPs in general and ZnO NPs in particular.
The present study offers an alternative method for green synthesis of the formation of two types of nanoparticles (NPs). These NPs, titanium oxide and silver NPs (TiO2 and Ag NPs, respectively), were obtained from the amalgamation of intracellular extract of a wild mushroom, Fomes fomentarius, with aqueous solutions of titanium isopropoxide and silver nitrate, respectively. F. fomentarius was identified phenotypically and by 18S ribosomal RNA gene sequencing (Gene accession no: MK635351). The biosynthesis of TiO2 and Ag NPs was studied and characterized by X-ray diffraction (XRD), diffuse reflectance UV-Visible spectroscopy (DR-UV), fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and transmission electron microscope (TEM). Success was achieved in obtaining NPs of differing sizes and shapes. The antibacterial and anticancer activity of the NPs was significant with morphological damage being caused by both, although Ag NPs (10–20 nm) were found to have profound effects on bacterial and cancer cells in comparison to TiO2 NPs (100–120 nm). These metal NPs, synthesized using wild mushrooms, hold a great potential in biomedicinedue to an effective enzyme combination, which permits them to modify different chemical compounds to less toxic forms, which is required for ecofriendly and safe biomaterials.
A b s t r a c tNon-specific and often misleading clinical presentation of active brucellosis has made it a diagnostic puzzle for treating physicians. Clinicians rely greatly on the detection of IgG and IgM anti-Brucella antibodies by ELISA. Different patterns of positivity have been observed for IgG and IgM anti-Brucella antibodies in different cases, which further increases the risk of an erroneous diagnosis. Detailed herein is our two-years data with varied Brucella serology patterns and their clinical interpretation. Between January 2015 to December 2017, 1102 samples were processed in the Immunology Laboratory of KFHU for Brucella serology. 68 samples were positive for both IgG and IgM, 28 samples were positive for IgG and negative for IgM while 15 samples were positive for IgM and negative for IgG antibodies against Brucella. Electronic medical records, history of exposure, signs, symptoms, laboratory data, and the final diagnosis were recorded for all these patients. None of the patients with only positive IgM antibodies was finally diagnosed with brucellosis, while a diagnosis of brucellosis was established for only one patient with IgG antibodies positive in his serum. All the double-positive (IgG-and IgM-positive) serology patterns were diagnosed as having brucellosis. We concluded that determination of single IgM or IgG anti-Brucella-antibodies by ELISA could both be considered as definite and should ideally be interpreted in the context of appropriate clinical scenario and confirmation by other laboratory assays. K e y w o r d s: Brucella-specific IgG, Brucella-specific IgM, Brucellosis, ELISA for Brucella
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