Rapid and Highly Sensitive Detection of Target DNA Related to COVID-19 Virus With a Fluorescent Bio-conjugated Probe via a FRET Mechanism.

A novel cyanine 3 (Cy3)-based bio-conjugated sensor has been developed to detect target DNA or extracted RNA from COVID -19 samples using the fluorescence resonance energy transfer (FRET) experiment. A special sequence of the COVID -19 genome was selected as a complementary DNA (target DNA) part. The opposite chain of this target sequence was designed in 2 parts; one part was attached to the Cy3 organic dye (capture DNA or Cy3- DNA), and the other part was attached to the BHQ2 molecule (quencher DNA or BHQ2- DNA). The Cy3 molecule acts as a donor pair, and BHQ2 acts as an acceptor pair in the FRET experiment. The capture DNA and quencher DNA can form a sandwiched complex in the presence of target DNA. The formation of the entitled sandwiched hybrid causes the decrement of emission intensity of the Cy3 donor in bio-conjugated Cy3-DNA via energy transfer from Cy3 (as a donor) to BHQ2 (as an acceptor). Indeed, in the presence of non-complementary DNA, the pairing of DNA strands does not occur, the FRET phenomenon does not exist, and therefore fluorescence intensity of Cy3 does not decrease. Moreover, this biosensor was successfully applied to analyze real samples containing extracted RNA of COVID -19 prepared for the reverse transcriptase-polymerase chain reaction (RT-PCR) test, and the results were promising.

Efficient and Versatile Application of Fluorescence DNA-Conjugated CdTe Quantum Dots Nanoprobe for Detection of a Specific Target DNA of SARS Cov-2 Virus.

Regarding the outbreak of the SARS Cov-2 virus pandemic worldwide, it seems necessary to provide new diagnostic methods to combat the virus. A fluorescence CdTe quantum dots-DNA (QDs-DNA) nanosensor was prepared for efficient detection of a specific target complementary DNA or RNA from the SARS Cov-2 virus using FRET experiment via forming a classic "sandwich" structure. The sequence of the complementary DNA (target DNA) is planned based on a substantial part of the SARS Cov-2 virus genome, and oligonucleotides of QDs-DNA nanoprobe are designed to complement it. The water-soluble CdTe QDs-DNA was prepared by replacing the thioglycolic acid (TGA) on the surface of QDs with capture DNA (thiolated DNA) through a ligand-exchange method. Subsequently, with the addition of complementary (target DNA) and quencher DNA (BHQ2-labeled DNA) into the QDs-DNA conjugates, sandwiched hybrids were formed. The resulting assembly brings the BHQ2-labeled DNA (as the acceptor), and the QDs (as the donor) into proximity, leading to quenching of fluorescence emission from the donor QDs through the FRET mechanism. In other words, a simple, highly sensitive, selective, and rapid approach was introduced to detect complementary DNA sequence from a specific part of the SARS Cov-2 virus genome with a detection limit of 2.52 × 10-9 mol L-1. Furthermore, the planned nanosensor was well used for the detection of RNA from SARS Cov-2 viruses in real samples with satisfactory analytical results, and the outcomes were compared with RT-PCR (Reverse Transcription Polymerase Chain Reaction) as the well-known standard method.

Novel CMC-CdTe / ZnS QDs Nanosensor for the Detection of Anticancer Drug Epirubicin.

Epirubicin (EPI) is one of the standard anticancer drugs that apply for various cancers treatment. However, the accumulation of EPI in the human body can be highly toxic, and it causes inevitable harm to organs. As a result, the evaluation of low concentrations of this drug in body samples requires sensitive, rapid, and accurate analysis methods. The fluorescence method is an efficient way in comparison of the traditional methods such as liquid chromatography, capillary electrophoresis, and electrochemical methods. Herein, we synthesized a novel fluorescence nanosensor named CMC-CdTe/ZnS based on using quantum dots (QDs). The structure of the prepared nanosensor is confirmed by different analysis methods such as FT-IR, TGA, and TEM. Besides that, the fluorescence intensity response of CMC-CdTe/ZnS QDs in the presence of Epirubicin drug is investigated. Based on obtained results, not only this nanosensor developed, but also the fluorescence quenching was explained by the typical Stern-Volmer equation. The best linear quenching equation for entitled nanosensor in the presence of Epirubicin is F0/F = 0.0346Q + 1.08 (R2 = 0.99), and the detection limit of Epirubicin is around 0.04 × 10-6 mol/L at 25 °C. All of the results display that this method could be reliable and suitable approach for determination of Epirubicin in commercial samples as well.

A 3-year study of Candida infections among patients with malignancy: etiologic agents and antifungal susceptibility profile.

Objective: Opportunistic fungal infections by Candida species arise among cancer patients due to the weakened immune system following extensive chemotherapy. Prophylaxis with antifungal agents have developed the resistance of Candida spp. to antifungals. Accurate identification of yeasts and susceptibility patterns are main concerns that can directly effect on the treatment of patients. Methods: Over a period of three years, 325 cancer patients suspected to Candida infections were included in the current investigation. The clinical isolates were molecularly identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). All strains, were examined for in vitro susceptibility to the amphotericin B, itraconazole, fluconazole, and anidulafungin according to the CLSI M27 document. Results: Seventy-four cancer patients had Candida infections (22.7%). Candida albicans was the most common species (83.8%). Antifungal susceptibility results indicated that 100% of the Candida isolates were sensitive to amphotericin B; however, 17.6%, 9.4%, and 5.4% of clinical isolates were resistant to anidulafungin, fluconazole, and itraconazole, respectively. Conclusion: The findings of the present work shows a warning increase in resistance to echinocandins. Since all fluconazole resistance isolates were obtained from candidemia, we recommend amphotericin B as the first line therapy for this potentially fatal infection.

Capability of novel fluorescence DNA-conjugated CdTe/ZnS quantum dots nanoprobe for COVID-19 sensing.

Urgent identification of COVID-19 in infected patients is highly important nowadays. Förster or fluorescence resonance energy transfer (FRET) is a powerful and sensitive method for nanosensing applications, and quantum dots are essential materials in FRET-based nanosensors. The QDs are conjugated to DNA or RNA and used in many applications. Therefore, in the present study, novel fluorescence DNA-conjugated CdTe/ZnS quantum dots nanoprobe designed for detection of Covid-19 after extracting their RNA from saliva of hesitant people. For achieving this purpose, the water-soluble CdTe/ZnS QDs-DNA prepared via replacing the thioglycolic acid (TGA) on the surface of QDs with capture DNA (thiolated DNA) throw a ligand-exchange method. Subsequently, by adding the different concentrations of complementary (target DNA) in a mixture of quencher DNA (BHQ2-labeled DNA) and the QDs-DNA conjugates at different conditions, sandwiched hybrids were formed. The results showed that the fluorescence intensity was decreased with increasing the concentration of target DNA (as a positive control). The linear equation and regression (Y = 40.302 X  + 1 and R2 = 0.98) were obtained by using the Stern-Volmer relationship. The Limit of detection (LOD) was determined 0.000823 µM. The achieved results well confirm the outcomes of the RT-PCR method in real samples.