Clinical Samples
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2021 ◽  
Vol 18 (1) ◽  
Kiran K. Mangalaparthi ◽  
Sandip Chavan ◽  
Anil K. Madugundu ◽  
Santosh Renuse ◽  
Patrick M. Vanderboom ◽  

AbstractSARS-CoV-2, a novel human coronavirus, has created a global disease burden infecting > 100 million humans in just over a year. RT-PCR is currently the predominant method of diagnosing this viral infection although a variety of tests to detect viral antigens have also been developed. In this study, we adopted a SISCAPA-based enrichment approach using anti-peptide antibodies generated against peptides from the nucleocapsid protein of SARS-CoV-2. We developed a targeted workflow in which nasopharyngeal swab samples were digested followed by enrichment of viral peptides using the anti-peptide antibodies and targeted parallel reaction monitoring (PRM) analysis using a high-resolution mass spectrometer. This workflow was applied to 41 RT-PCR-confirmed clinical SARS-CoV-2 positive nasopharyngeal swab samples and 30 negative samples. The workflow employed was highly specific as none of the target peptides were detected in negative samples. Further, the detected peptides showed a positive correlation with the viral loads as measured by RT-PCR Ct values. The SISCAPA-based platform described in the current study can serve as an alternative method for SARS-CoV-2 viral detection and can also be applied for detecting other microbial pathogens directly from clinical samples.

2021 ◽  
Vol 499 ◽  
pp. 113160
Sasha E. Larsen ◽  
Bryan J. Berube ◽  
Tiffany Pecor ◽  
Evan Cross ◽  
Bryan P. Brown ◽  

2021 ◽  
Qihua XU ◽  
Zi-Yi Wang

Abstract Proliferative vitreoretinopathy (PVR) is the main reason for the failure of retinal detachment surgeryEpithelial-mesenchymal transition (EMT) induced by transforming growth factor (TGF-β2) plays an important role in the development of PVR.Artesunate has been widely studied in the treatment of ophthalmic diseases because of its antioxidant, anti-inflammatory, anti-apoptosis and anti-proliferation effects.The purpose of this study was to investigate the effect of artesunate on EMT induced by TGF-β2 in ARPE-19 cells and its effect on PVR processWe found that artesunate can inhibit the proliferation of ARPE-19 cells after EMT transformation, inhibit the contraction of ARPE-19 cells after EMT transformation, and inhibit the autocrine of TGF-β2 in ARPE-19 cellsWe also found that the contents of Smad3 and p-smad3 in clinical samples increased,Artesunate can inhibit the contents of Smad3 and p-smad3 in ARPE-19 cells induced by TGF-β2Artesunate can inhibit the occurrence and development of PVR diseases in vivoTo sum up, artesunate can inhibit the occurrence and development of PVR diseases by inhibiting the EMT process of ARPE-19 cells.

2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Amaka Marian Awanye ◽  
Chidozie Ngozi Ibezim ◽  
Catherine Nonyelum Stanley ◽  
Hannah Onah ◽  
Iheanyi Omezurike Okonko ◽  

2021 ◽  
Vol 11 (1) ◽  
Kristína Boršová ◽  
Evan D. Paul ◽  
Viera Kováčová ◽  
Monika Radvánszka ◽  
Roman Hajdu ◽  

AbstractThe emergence of a novel SARS-CoV-2 B.1.1.7 variant sparked global alarm due to increased transmissibility, mortality, and uncertainty about vaccine efficacy, thus accelerating efforts to detect and track the variant. Current approaches to detect B.1.1.7 include sequencing and RT-qPCR tests containing a target assay that fails or results in reduced sensitivity towards the B.1.1.7 variant. Since many countries lack genomic surveillance programs and failed assays detect unrelated variants containing similar mutations as B.1.1.7, we used allele-specific PCR, and judicious placement of LNA-modified nucleotides to develop an RT-qPCR test that accurately and rapidly differentiates B.1.1.7 from other SARS-CoV-2 variants. We validated the test on 106 clinical samples with lineage status confirmed by sequencing and conducted a country-wide surveillance study of B.1.1.7 prevalence in Slovakia. Our multiplexed RT-qPCR test showed 97% clinical sensitivity and retesting 6,886 SARS-CoV-2 positive samples obtained during three campaigns performed within one month, revealed pervasive spread of B.1.1.7 with an average prevalence of 82%. Labs can easily implement this test to rapidly scale B.1.1.7 surveillance efforts and it is particularly useful in countries with high prevalence of variants possessing only the ΔH69/ΔV70 deletion because current strategies using target failure assays incorrectly identify these as putative B.1.1.7 variants.

2021 ◽  
Vol In Press (In Press) ◽  
Folasade Muibat Adeyemi ◽  
Nana-Aishat Yusuf ◽  
Rashidat Ronke Adeboye ◽  
Omotayo Opemipo Oyedara

Background: The virulence factors of enterococci play a major role in the pathogenicity of enterococcal strains. Objectives: This study aimed to evaluate virulence factors and detect selected virulence and resistance genes in vancomycin-resistant Enterococcus (VRE) from clinical samples from southwest Nigeria. Methods: The VRE isolates (n = 85) recovered from clinical samples were characterized using conventional microbiology techniques, and molecular identification was made with ddlE primers. Phenotypic screening for five virulence determinants and detection of virulence and resistance genes using a polymerase chain reaction were carried out. Results: Phenotypic identification revealed 61 Enterococcus faecium and 24 Enterococcus faecalis. All the isolates hydrolyzed bile. Moreover, 88.2% of the isolates produced biofilm; however, 72.9% of the isolates produced gelatinase enzyme. Altogether, six isolates (7%) produced all five virulence factors. The least virulence factor expressed by the two species E. faecium and E. faecalis was DNase at 21.3% and 29.2% followed by cytolysin at 27.9% and 41.7%, respectively. Only 25 isolates (29.4%), including 23 E. faecium (37.7%) and only 2 (8.3%) E. faecalis isolates, revealed bands with molecular identification. Additionally, VRE isolates showed bands for asa1 (16%); only 1 (4%) isolate had the hyl gene and vanB gene, respectively. Conclusions: The absence of vanA and low detection of vanB resistance genes suggest the possible presence of other van types and emphasizes the need for further investigations on the incidence of other van genes using molecular screening methods in enterococci isolates in Nigeria for surveillance purposes. Moreover, the low occurrence of virulence genes implies that there might be other mediators of pathogenicity involved in Enterococcus virulence traits.

Yara Elahi ◽  
Jamileh Nowroozi ◽  
Ramin Mazaheri Nezhad Fard

Background and Objectives: In recent decades, enterococcal resistance to antimicrobials has greatly increased. Further- more, these chemicals include several side effects on the patients. Since no reports are available of the bacteriophages' effects on eukaryotic cells, they can be good solutions for multidrug-resistant bacterial problems. Therefore, the major aim of this study was to isolate bacteriophages from wastewaters on clinical antibiotic-resistant enterococci. Materials and Methods: Clinical bacteria were isolated, then enterococcal isolates were identified using different methods. The antibiotic resistance scheme of the enterococcal isolates was assessed. The bacterial isolates were exposed to wastewa- ter samples containing potential bacteriophages. Technically, isolated bacteriophages were studied by electron microscopy. Results: Isolated bacteria were verified as Enterococcus faecium. Results showed that bacteriophages could easily be isolat- ed from wastewater sources. The isolated bacteriophages were effective on E. faecium as well as Streptococcus dysgalactiae. Furthermore, these bacteriophages were challenged with five other bacteria (ATCC) with no visible effects. In general, the isolated bacteriophages belonged to the Myoviridae, Siphoviridae, and Inoviridae families. Conclusion: Further studies on bacteriophages and their efficacy on enterococcal strains could increase the treatment possi- bility of enterococcal infections. Due to these bacteriophages' effects on Streptococcus strains, bacteriophages may be used to treat streptococcal infections as well.

M. Anjaneya Swamy ◽  
Jagannath D. Andhale

Background: Since there is a significant rise in resistant bacteria to different antimicrobial agents, there is a need to study the resistance pattern of different isolates from different clinical samples for effective use of available antimicrobials by clinicians. The aim of the present study was to detect the resistance pattern of various antimicrobials against different clinical isolates in hospitalised patients in out setting.Methods: This is a retrospective study involving the collection of the data from the records of microbiology laboratory. All clinical specimens were processed as per standard microbiological procedures. Antibiotic susceptibility testing was performed by Kirby Bauer disc diffusion method on Mueller Hinton agar plate as per CLSI guidelines.Results: A total of 153 isolates were recovered from 219 clinical samples accounting for 69.86% of total positivity. Which includes gram negative bacilli 107/153 (69.93%) gram positive cocci 36/153 (23.53%) and yeast 10/153 (6.54%). Among the total isolates gram negative bacilli account for major number of isolates 69.93% followed by gram positive cocci 23.53% and yeast 6.54%. Gram positive cocci and gram-negative bacilli showed a significant level of antimicrobial resistance. Nitrofurantoin is highly effective against urinary isolates of Escherichia coli. vancomycin and linezolid are most effective antimicrobials against gram positive cocci. Among gram negative bacilli meropenem and amikacin are most effective antimicrobials. Statistical significance of occurrence of Escherichia coli as predominant isolate as compared to other isolates were analysed by chi square test by using GraphPad online calculator. A p value<0.001 was obtained.Conclusions: Significant rise in antimicrobial resistant pathogens were observed. Local antimicrobial policy should be developed for effective selection of available antimicrobials which are the need of the day to reduce the burden of diseases on global health care system.

Cancers ◽  
2021 ◽  
Vol 13 (20) ◽  
pp. 5051
Inti Peredo-Harvey ◽  
Afsar Rahbar ◽  
Cecilia Söderberg-Nauclér

Glioblastoma is a malignant brain tumor with a dismal prognosis. The standard treatment has not changed in the past 15 years as clinical trials of new treatment protocols have failed. A high prevalence of the human cytomegalovirus (HCMV) in glioblastomas was first reported in 2002. The virus was found only in the tumor and not in the surrounding healthy brain tissue. Many groups have confirmed the presence of the HCMV in glioblastomas, but others could not. To resolve this discrepancy, we systematically reviewed 645 articles identified in different databases. Of these, 81 studies included results from 247 analyses of 9444 clinical samples (7024 tumor samples and 2420 blood samples) by different techniques, and 81 articles included 191 studies that identified the HCMV in 2529 tumor samples (36% of all tumor samples). HCMV proteins were often detected, whereas HCMV nucleic acids were not reliably detected by PCR methods. Optimized immunohistochemical techniques identified the virus in 1391 (84,2%) of 1653 samples. These data suggest that the HCMV is highly prevalent in glioblastomas and that optimized immunohistochemistry techniques are required to detect it.

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