Blood is the most interesting tissue for molecular analysis in a wide range of diseases since it provides accumulative information of the whole body state. Transcriptomic analysis of the blood is one of the most favorable omics approaches for clinical and research purposes. However, blood sample preparation for RNA analyses is highly diverse encompassing specific methodology and application. In this study, we review the common sample preparation maneuvers including RNA handling techniques, sample collection and stabilization, blood cell sorting, extracellular vesicle enrichment, RNA extraction, globin/ribosomal RNA reduction, and RNA concentration and quality assessment. These techniques are the most prevalent procedures of blood sample preparation in RNA transcriptomic analyses. The methodologies, technical challenges of the current approaches, and latest advances in the field are discussed in detail.
We developed a microchip device using surface acoustic waves (SAW) and sharp‐edge glass microparticles to rapidly lyse low‐level cell samples. This microchip features a 13‐finger pair interdigital transducer (IDT) with a 30‐degree focused angle, creating high‐intensity acoustic beams converging 6 mm away at a 16 MHz frequency. Cell lysis is achieved through centrifugal forces acting on Candida albicans cells and glass particles within the focal area. To optimize this SAW‐induced streaming, we conducted 42 pilot experiments, varying electrical power, droplet volume, glass particle size, concentration, and lysis time, resulting in optimal conditions: an electrical signal of 2.5 W, a 20 μL sample volume, glass particle size below 10 μm, concentration of 0.2 μg, and a 5‐min lysis period. We successfully amplified DNA target fragments directly from the lysate, demonstrating an efficient microchip‐based cell lysis method. When combined with an isothermal amplification technique, this technology holds promise for rapid point‐of‐care (POC) applications.
15 June 2019 in Journal of Microbiology and Infectious Diseases
Objective: Human Cytomegalovirus (CMV) is a member of the Herpesviridae family, with the ability to establish a long-lived latent infection. CMV infection causes problems in immunocompromised hosts undergoing organ and stem cell transplantation. The prevalence of CMV in adults varies in different geographic regions. The purpose of this study was to assess the prevalence of CMV in the adult population of Afghanistan, which did not have epidemiologic information for CMV infection. Methods: A total of 500 adults residing in main regional provinces of Afghanistan including Nangarhar, Herat, Mazar-i Sharif, Kandahar and Kabul in the age range of 25-70 years old were randomly selected to include in the study. Among the participants, 263 (52.6%) were female and 237 (47.4%) were male. The samples were tested for the presence of CMV IgM and IgG antibodies using chemiluminescence immunoassay on the Abbott Architect automated platform. Results: The seropositivity of CMV was found 99.79 % in Afghanistan. There were no significant differences in the prevalence CMV infection among the five regions. The seropositivity anti-CMV IgG positive rate in Kandahar, Kabul, Nangarhar and, Herat was determined as 100%. The CMV IgG prevalence was 98.99% in Mazar-i Sharif. Anti CMV IgM was found 1.24% in Afghanistan. Conclusions: Our study shown that the seroprevalence is high in Afghanistan. Because of the high frequency of seropositivity in general population in Afghanistan, the approaches for preventing CMV reactivation need to be developed. J Microbiol Infect Dis 2019; 9(2): 78-82.