scholarly journals SARS-CoV-2 detection in primary thyroid sarcoma: coincidence or interaction?

2022 ◽  
Author(s):  
M. L. Tanda ◽  
S. Ippolito ◽  
D. Gallo ◽  
A. Baj ◽  
F. Novazzi ◽  
...  
Keyword(s):  
Scientific Literature ◽  
Needle Aspiration ◽  
Whole Genome Sequence ◽  
Ultrastructural Examination ◽  
Functional Studies ◽  
Cytoplasmic Vacuoles ◽  
Mouse Double Minute ◽  
Viral Particles ◽  
Polymerase Chain ◽  
Thyroid Dysfunctions

Abstract Introduction Thyroid dysfunctions associated with SARS-CoV-2 are emerging in scientific literature. During the second COVID-19 epidemic spread, we evaluated a patient with the suspect of subacute thyroiditis. Methods and results Specimen from fine-needle aspiration of a hypoechoic undefined area was analyzed for cytology and for SARS-CoV-2 detection. SARS-CoV-2 was retrieved by real-time polymerase chain reaction on the cytologic sample, which was then cultured on Vero E6 cells and demonstrated to be cytopathic. Whole-genome sequence was deposited. Histological exam diagnosed a rare case of primary thyroid sarcoma with diffuse and strong expression of mouse double minute 2 homolog (MDM2) oncoprotein. Ultrastructural examination confirmed, in several neoplastic cells, the presence of viral particles in cytoplasmic vacuoles. Conclusions In our hypothesis, SARS-CoV-2 and sarcoma coexistence could represent a synergistic interplay, ultimately favoring both viral persistence and tumor proliferation: the overexpression of MDM2 in tumor cells might have generated a favorable immunological niche for SARS-CoV-2 localization and, in turn, SARS-CoV-2 could have favored tumor growth by inducing MDM2-mediated p53 downregulation. Functional studies are needed to confirm this suggestive pathway.

scholarly journals Development of RT-PCR Based Diagnosis of SARS-CoV-2

2021 ◽  
Author(s):  
Rutuja Sunil Patankar ◽  
Vasudeo Pandharinath Zambare
Keyword(s):  
World Wide ◽  
Method Development ◽  
Vaccine Development ◽  
Whole Genome Sequence ◽  
Molecular Technique ◽  
Rt Pcr ◽  
Global Priority ◽  
Diagnosis Method ◽  
Pcr Method ◽  
Polymerase Chain

In the 2020, COVID-19 pandemic disease created an havoc situation world widely and mainly caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). It has been challenging task for researchers, scientists and medico-pharmaceutical organisations to find out rapid and reliable diagnosis methods. Among the all testing services, a Reverse Transcription Polymerase Chain Reaction (RT-PCR) is the more accurate, rapid and authenticated molecular technique used for most of the diagnosis of major diseases. It has been a global priority to fix the rapid diagnosis method to combat against the pandemic COVID-19. Thus, the present chapter mainly focussing on the progress of RT-PCR method development though various processes of data collection on isolation of whole genome sequence, its primer and method designing. In this scenario, India suddenly become the global leader for vaccine development and hence the challenges and RT-PCR kit development in India and rest of the world has been be discussed. World wide many Government and private agencies and industries have taken an initiative for diagnosis of SARS-CoV-2 hence this chapter also summarised the scope of RT-PCR to combat pandemic situation in future.

scholarly journals Are There 1031 Virus Particles on Earth, or More, or Fewer?

2020 ◽  
Vol 202 (9) ◽  
Author(s):  
A. R. Mushegian
Keyword(s):  
Scientific Literature ◽  
General Interest ◽  
Virus Particles ◽  
Know How ◽  
Link Type ◽  
Best Estimate ◽  
Viral Particles ◽  
History Of ◽  
New Evidence

ABSTRACT The number of virus particles on Earth is frequently reported in the scientific literature and in general-interest publications as being on the order of 1031, with some confusion about whether this is a high or low estimate. This number is often given without a source, although it should be attributed to a paper by Hendrix et al. published in 1999 (R. W. Hendrix, M. C. Smith, R. N. Burns, M. E. Ford, and G. F. Hatfull, Proc Natl Acad Sci U S A 96:2192–2197, 1999, https://doi.org/10.1073/pnas.96.5.2192). As with any oft-repeated statistic, it is informative to know how it has been derived and whether it should be revised in the light of new evidence. I review the history of the 1031 estimate and use more recent assessments of the number of bacterial and viral particles in various habitats to conclude that the best estimate of the number of virus particles on Earth (“the Hendrix product”) remains close to 1031 and is unlikely to be either much less or much more than that.

scholarly journals A One-Tube Polymerase Chain Reaction Protocol Demonstrates CC Chemokine Overexpression in Graves’ Disease Glands

1999 ◽  
Vol 84 (8) ◽  
pp. 2873-2882
Author(s):  
Yaqoub Ashhab ◽  
Orlando Dominguez ◽  
Mireia Sospedra ◽  
Carme Roura-Mir ◽  
Anna Lucas-Martín ◽  
...  
Keyword(s):  
Human Leukocyte ◽  
Needle Aspiration ◽  
Graves Disease ◽  
Tissue Samples ◽  
Autoimmune Thyroid ◽  
Leukocyte Antigen ◽  
Cc Chemokine ◽  
Inflammatory Protein ◽  
Cc Chemokines ◽  
Polymerase Chain

An adaptation of mixed oligonucleotide primed amplification of complementary DNA to detect the profile of CC chemokines in biological samples is presented. By introducing normalization, two correction coefficients, performing a single amplification reaction, and five parallel hybridizations, intrasample and intersample comparisons can be reliably made. This protocol of single tube PCR CC chemokine profiling was applied to tissue samples from an autoimmune thyroid condition, Graves’ disease, and from a nonautoimmune condition, multinodular goiter. Results demonstrate overexpression of CC chemokines in Graves’ disease, statistically significant for macrophage inflammatory protein-1α and -1β, which correlated with the aberrant human leukocyte antigen class II expression by thyrocytes, as assessed by flow cytometry. Overexpression of CC chemokines probably plays a major role in determining the characteristics of the lymphocytes migrating to the thyroid gland and influences the course of the disease. The study of chemokine profile should be more informative than the study of isolated chemokines and cytokines, and as it can be applied to fine needle aspiration biopsies, it may be useful to clinical research.

SOCS1 and SHP1 hypermethylation in multiple myeloma: implications for epigenetic activation of the Jak/STAT pathway

Blood ◽  
2004 ◽  
Vol 103 (12) ◽  
pp. 4630-4635 ◽  
Author(s):  
Chor-Sang Chim ◽  
Tsz-Kin Fung ◽  
Wai-Chung Cheung ◽  
Raymond Liang ◽  
Yok-Lam Kwong
Keyword(s):  
Polymerase Chain Reaction ◽  
Promoter Hypermethylation ◽  
Janus Kinase ◽  
Chain Reaction ◽  
Functional Studies ◽  
Stat3 Phosphorylation ◽  
Translation Start ◽  
Polymerase Chain ◽  
Phosphorylated Stat3 ◽  
Stat Pathway

Abstract SOCS1 and SHP1 negatively regulate the Janus kinase/signal transducer and activator of transcription (Jak/STAT) signaling pathway. The role of promoter hypermethylation leading to epigenetic inactivation of SOCS1 and SHP1 in myeloma was investigated. The methylation-specific polymerase chain reaction (MSP) was used to define SOCS1 and SHP1 methylation in 34 diagnostic myeloma samples. For SOCS1, MSP primers 3′ to the translation start site were unreliable and gave positive results in normal controls. However, primers in the 5′ promoter region were specific, although no myeloma samples showed methylation. For SHP1, 27 of 34 (79.4%) myeloma samples showed SHP1 hypermethylation. The biologic significance of SHP1 methylation was investigated in the U266 human myeloma line. U266 contained completely methylated SHP1. Furthermore, there was constitutive STAT3 phosphorylation. Treatment with 5-azacytidine led to progressive demethylation of SHP1 on days 2 to 5, with consequent increasing reexpression of SHP1 as shown by reverse transcription-polymerase chain reaction (RT-PCR). Concomitant with increasing SHP1, a parallel down-regulation of phosphorylated STAT3 occurred, so that by day 5 phosphorylated STAT3 was barely detectable. The overall survivals of patients with and without SHP1 methylation were similar. SHP1 methylation leading to epigenetic activation of the Jak/STAT pathway might have a tentative role in the pathogenesis of myeloma, which should be further confirmed by functional studies in primary myeloma samples. (Blood. 2004;103:4630-4635)

scholarly journals Multiplex PCR for detection of MCR genes in clinical fecal samples

2021 ◽  
Vol 269 ◽  
pp. 01019
Author(s):  
Qiumei Xiang ◽  
Shuanglan Hu ◽  
Yuebin Ke ◽  
Shuangfang Hu
Keyword(s):  
Clinical Laboratory ◽  
Sequence Data ◽  
Rapid Identification ◽  
Whole Genome Sequence ◽  
Epidemiological Surveillance ◽  
Universal Primers ◽  
Colistin Resistance ◽  
Pcr Assay ◽  
Polymerase Chain ◽  
Higher Sensitivity

Plasmid-mediated colistin-resistance genes have been reported worldwide in recent years. A multiplex polymerase chain reaction (Multi-PCR) protocol was developed to detect transferable colistinresistance genes (mcr-1 to mcr-6) in Enterobacteria for clinical laboratory purposes.The authors first designed six new primer pairs to amplify mcr-1 to mcr-6 gene products to achieve stepwise separation of amplicons between 87 to 216 bp,then divided these primers into two subgroups with the assistance of a pair of universal primers for the detection of currently described mcr genes and their variants in Enterobacteria. The protocol was validated by testing 29 clinical isolates of Escherichia coli of human origin, each well characterised and prospectively validated. The Multi-PCR assay showed full concordance with whole-genome sequence data and displayed higher sensitivity and 100% specificity. The assay could detect all variants of the various mcr alleles described. It was able to detect mcr-3 and mcr-4 as singletons or in combination. This type of test is critical for the epidemiological surveillance of plasmid-encoded resistance in limited resources conditions, and this method allows rapid identification of mcr-positive bacteria and overcomes the challenges of phenotypic detection of colistin resistance.

scholarly journals An optimised protocol for detection of SARS-CoV-2 in stool

2021 ◽  
Author(s):  
T Li ◽  
E Garcia-Gutierrez ◽  
J Scadden ◽  
J Davies ◽  
C Hutchins ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Rna Extraction ◽  
Extraction Procedure ◽  
Clinical Practices ◽  
Faecal Microbiota ◽  
Chain Reaction ◽  
Oral Transmission ◽  
Viral Particles ◽  
Stool Samples ◽  
Polymerase Chain

AbstractAimSARS-CoV-2 has been detected in stool samples of COVID-19 patients, with potential implications for faecal-oral transmission. Compared to swab samples, the complexity of the stool matrix poses a challenge in the detection of the virus that has not yet been solved. The aim of this study was to establish a sensitive and reliable method for detecting SARS-CoV-2 in stool samples.MethodsStool samples from individuals free of SARS-CoV-2 were homogenised in saline buffer and spiked with a known titre of inactivated virus ranging from 50 to 750 viral particles per 100 mg stool. Debris was removed via centrifugation and supernatants were concentrated by ultrafiltration. RNA was then extracted from the concentrated material using a commercial kit and SARS-CoV-2 was detected via real-time reverse-transcription polymerase chain reaction (RT-qPCR) using the CDC primers and probes.ResultsThe RNA extraction procedure we used allowed the detection of SARS-CoV-2 via RT-qPCR in most of the stool samples tested. We could detect as few as 50 viral particles per 100 mg of stool. However, high variability was observed across samples at low viral titres. The primer set targeting the N1 region provided more reliable and precise results and for this primer set our method had a limit of detection of 1 viral particle per mg of stool.ConclusionsHere we describe a sensitive method for detecting SARS-CoV-2 in stool samples. This method can be used to establish the persistence of SARS-CoV-2 in stool and ensure the safety of clinical practices such as faecal microbiota transplant (FMT).

scholarly journals Problems and Solution to Diagnose Extrapulmonary Tuberculosis in Central Region of Nepal

Med Phoenix ◽  
2017 ◽  
Vol 1 (1) ◽  
pp. 41-43
Author(s):  
Ravi Shankar Gupta ◽  
Tarannum Khatun ◽  
Akhtar Alam Ansari ◽  
Amrullah Shidiki ◽  
Dipak Bhargava ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Correct Diagnosis ◽  
Extrapulmonary Tuberculosis ◽  
Needle Aspiration ◽  
Retropharyngeal Abscess ◽  
Chain Reaction ◽  
Specificity And Sensitivity ◽  
Polymerase Chain ◽  
Needle Aspiration Cytology

Extrapulmonary Tuberculosis is high, challenging the clinicians to make correct diagnosis. Microscopy, culture and fine needle aspiration cytology have their limitations in regard to specificity and sensitivity. In this report, polymerase chain reaction is used for detecting and distinguishing Extrapulmonary Tuberculosis. A case of retropharyngeal abscess was selected from which pus was collected which was negative for microscopy and culture in routine microbiology as well as mycobacteriology. Cytopathological examination was also negative. Polymerase chain reaction was applied to detect Mycobacterium tuberculosis specific IS6110 gene. The patients responded with anti-tuberculosis treatment well. Polymerase chain reaction was introduced for diagnosis of Extrapulmonary Tuberculosis since it can be done within hours, monitor therapy and also differentiate Mycobacterium tuberculosis from other Mycobacterial species.MED Phoenix Volume (1), Issue (1) July 2016, page: 41-43

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