Detection of Viruses by Multiplex Real-Time Polymerase Chain Reaction in Bronchoalveolar Lavage Fluid of Patients with Nonresponding Community-Acquired Pneumonia
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Detection of Viruses by Multiplex Real-Time Polymerase Chain Reaction in Bronchoalveolar Lavage Fluid of Patients with Nonresponding Community-Acquired Pneumonia
Background: Non-responding pneumonia is responsible for the largest mortality of community-acquired pneumonia (CAP). However, until now, it is not clear whether viral infection plays an important role in the etiology of the non-corresponding cap and if there is a significant difference in the clinical characteristics between the viral non-respecting cap. respective.
Methods: From 2016 to 2019, patients with non-responding capsules have been recorded retrospectively in our study. All patients received bronchoalvolar washing (ball) and virus detection in the ball fluid by reaction of the polymerase multiplex chain (PCR) and clinical data, laboratories and radiographic laboratories were collected.
Results: A total of 43 patients were included. The median age was 62 and 65.1% of patients were men. Overall, 20 patients (46.5%) have been identified with viral infection. Of these viruses, the influenza virus (n = 8) and adenovirus (n = 7) have been detected more often, and other people included the herpes simplex virus, human enterovirus, cytomegalovirus The human coronavirus 229E, rhinovirus and the parainfluenza virus. Compared to the non-respective non-respective cap, only a soil opacity combined with the consolidation was a more common imaging manifestation in the non-corresponding viral cap. However, no obvious difference was found in the clinical and laboratory conclusions between the presence and absence of viral infections.
Conclusions: Viral infections were particularly common in adults with a non-corresponding cap. The soil opacity associated with the consolidation was a specific imaging manifestation for the non-corresponding viral cap, while clinical and laboratory data showed no obvious difference between the non-respective viral and non-respective cap.
The reaction of the quantitative polymerase chain of methylation-sensitive methylation allows rapid, precise and precise detection of the methylation state of the T cell area of the T cell (Treg) – specific demethylation in primary human rules
Human regulatory T cells (Tregs) have been involved in cancer immunotherapy and are also emerging cellular therapy for the treatment of several indications. Although Trega stability during ex vivo culture has improved, methods for evaluating Treg stability, such as bisulfite sequencing Sanger to determine the state of methylation of the demethylated region specific to the Treg (TSDR) remained unchanged. Bisulfite Sanger Sequencing is not only expensive and cumbersome to perform, it is incorrect because of relatively low read accounts. The sequencing of the next generation bisulfite, although more precise, is a less accessible method.
In this study, we describe the application of restriction enzymes sensitive to methylation (MSRE) and Quantitative PCR (QPCR) to determine the methylation status of the TSDR. Using known Tregs and Non-Tregs reports, we show that Msre-QPCR can distinguish the methylation status of the TSDR in cell populations containing a growing proportion of Tregs from 0 to 100%. In a comparison with values obtained from a bisulfite sequencing approach established to determine the methylation status of the TSDR, our MSRE-QPCR results were 5% on average for all samples with a high percentage (> 70% ) from Tregs, reinforcing that MSRE-QPCR can be completed in less time than other methods with the same level of precision.
The value of this assay was then demonstrated by quantifying the differences in treated Treg methylation status with and without rapamycin during an ex vivo expansion culture. Together, we show that our new msre-QPCR application to the TSDR is an optimal test for a precise assessment of TREG purity.
Detection of Viruses by Multiplex Real-Time Polymerase Chain Reaction in Bronchoalveolar Lavage Fluid of Patients with Nonresponding Community-Acquired Pneumonia
Analysis of extracellular bladder vesicle proteins using a digital polymerase chain reaction amplification
It is necessary for new analytical techniques to study the composition of simple extracellular vesicles (EV). These techniques are necessary to improve the understanding of the populations of EV heterogeneous in order to allow the identification of unique subpopulations and to allow detection of more than earlier and sensitive diseases. Due to the small size of EV and their low protein content, ultrachage sensitivity technologies are needed.
Here, a method of amplifying the reaction of the digital polymerase chain (IDDPCR) is described to allow multiplexed single-EV protein profiling. Antibody DNA conjugates are used to label EV, followed by a stochastic microfluidic incorporation of the single EV in the droplets. In situ PCR with fluorescent journalist probes converts and amplifies the barcode signal for subsequent reading by Goutteletelterie imaging. In these principle studies, it is shown that the analysis of the multiplex protein is possible in a single EV, opening the door for future analyzes. In this study, a poly-L-lysine magnetic pearl technique (PLL) of vancomycin (PLL) is developed for an insulation of gram-positive bacteria. The method combines a magnetic separation with a PCR (MPCR) multiplex dosage and gel electrophoresis for easy and fast detection of Bacillus cereus.
Description: DNA polymerase eta (POLH), is a protein that in humans is encoded by the POLH gene. This gene encodes a member of the Y family of specialized DNA polymerases. It copies undamaged DNA with a lower fidelity than other DNA-directed polymerases. However, it accurately replicates UV-damaged DNA; when thymine dimers are present, this polymerase inserts the complementary nucleotides in the newly synthesized DNA, thereby bypassing the lesion and suppressing the mutagenic effect of UV-induced DNA damage. This polymerase is thought to be involved in hypermutation during immunoglobulin class switch recombination. Mutations in this gene result in XPV, a variant type of xeroderma pigmentosum. Several transcript variants encoding different isoforms have been found for this gene.
Description: DNA polymerase iota is an enzyme that in humans is encoded by the POLI gene. The protein encoded by this gene is an error-prone DNA polymerase involved in DNA repair. The encoded protein promotes DNA synthesis across lesions in the template DNA, which other polymerases cannot do. The encoded polymerase inserts deoxynucleotides across lesions and then relies on DNA polymerase zeta to extend the nascent DNA strand to bypass the lesion.
Description: Polymerase (DNA directed), beta, also known as POLB, is an enzyme that, in humans, is encoded by the POLB gene. It is localized on 8p11.2. The protein encoded by this gene is a DNA polymerase involved in base excision and repair, also called gap-filling DNA synthesis. It is found that a truncated POLB is expressed in primary colorectal tumors and inhibits the normal repair function of wildtype POLB. The encoded protein, acting as a monomer, is normally found in the cytoplasm, but it translocates to the nucleus upon DNA damage. Several transcript variants of this gene exist, but the full-length nature of only one has been described to date. Additionally, human POLB forms a complex with and is methylated by PRMT6. In vitro, methylated POLB possesses significantly higher DNA polymerase activity when compared to that of unmodified enzyme. The increase in DNA polymerase activity upon methylation is due to the enhanced DNA binding and processivity of POLB.
Description: The protein encoded by this gene is a DNA polymerase involved in base excision and repair, also called gap-filling DNA synthesis. The encoded protein, acting as a monomer, is normally found in the cytoplasm, but it translocates to the nucleus upon DNA damage. [RefSeq]
Description: DNA polymerase lambda, also known as POLL, is a protein found in eukaryotes. In humans, it is encoded by the POLLA gene. POLL has 5-prime-deoxyribose-5-phosphate lyase activity and strand-displacement synthesis activity on gapped DNA substrates, suggesting that POLL participates in short- and long-patch base excision repair. POLL is widely expressed, with abundant expression in pachytene spermatocytes of testis and in ovary.
Vancomycin has been used as a molecular ligand between the MB and the d-alanyl-d-alanine boundaries on the surface of the cell wall of B. Cereus. The PLL served as a flexible molecular attachment between the MB and the van that reduced the obstacle to maintaining the biological activity of the van. The MB-PLL-VAN Capture nanoprobes have presented excellent capture and insulation efficiency for B. Cereus in milk matrix samples with interference from the complex food matrix. The following MPCR test showed a high specificity for the 4 target genes of B. Cereus, ENTFM, CESB, CER and 16S RNRA genes used to obtain effective genotyping and detection.
