Isolation of Neurocysticercosis-Related Antigens from a Genomic Phage Display Library of Taenia solium

2010 ◽  
Vol 15 (10) ◽  
pp. 1268-1273 ◽  
Author(s):  
Erik González ◽  
Yetzi Robles ◽  
Tzipe Govezensky ◽  
Raul J. Bobes ◽  
Goar Gevorkian ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Copy Number ◽  
Genomic Dna ◽  
Taenia Solium ◽  
Phage Display Library ◽  
Clinical Samples ◽  
Expression Library ◽  
Direct Identification ◽  
Antigen Discovery ◽  
Host Pathogen Interaction

In this study, the authors have generated a tapeworm Taenia solium genomic DNA expression library where foreign peptides/proteins were fused to N-termini of M13 cpVIII and expressed at a high copy number on the phage surface, and they showed that this library may be used in bioselection against antipathogen immune sera, allowing the identification of disease-related antigens recognizing antibodies present in clinical samples. They isolated 2 phage clones expressing T. solium–derived antigens specifically reacting with antibodies present in plasma and cerebrospinal fluid samples of neuroimaging-confirmed neurocysticercosis patients. The described antigen discovery strategy may be used for the direct identification of antigens useful for host-pathogen interaction studies as well as for the development of molecular vaccines and diagnostics.

scholarly journals Cytokine response in cerebrospinal fluid of meningitis patients and outcome associated with pneumococcal serotype

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Annelies Müller ◽  
Diana B. Schramm ◽  
Jackie Kleynhans ◽  
Linda de Gouveia ◽  
Susan Meiring ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Growth Factors ◽  
South African ◽  
Capsular Polysaccharide ◽  
Case Fatality ◽  
Bayesian Regression ◽  
Clinical Samples ◽  
Host Pathogen Interaction ◽  
Life Threatening ◽  
Pneumococcal Serotype

AbstractStreptococcus pneumoniae causes life-threatening meningitis. Its capsular polysaccharide determines the serotype and influences disease severity but the mechanism is largely unknown. Due to evidence of elevated cytokines levels in the meningeal inflammatory response, we measured 41 cytokines/chemokines and growth factors in cerebrospinal fluid (CSF) samples from 57 South African meningitis patients (collected in the period 2018–2019), with confirmed S. pneumoniae serotypes, using a multiplexed bead-based immunoassay. Based on multivariable Bayesian regression, using serotype 10A as a reference and after adjusting for HIV and age, we found IL-6 concentrations significantly lower in patients infected with serotypes 6D (undetectable) and 23A (1601 pg/ml), IL-8 concentrations significantly higher in those infected with 22A (40,459 pg/ml), 7F (32,400 pg/ml) and 15B/C (6845 pg/ml), and TNFα concentration significantly higher in those infected with serotype 18A (33,097 pg/ml). Although a relatively small number of clinical samples were available for this study and 28% of samples could not be assigned to a definitive serotype, our data suggests 15B/C worthy of monitoring during surveillance as it is associated with in-hospital case fatality and not included in the 13-valent polysaccharide conjugate vaccine, PCV13. Our data provides average CSF concentrations of a range of cytokines and growth factors for 18 different serotypes (14, 19F, 3, 6A, 7F, 19A, 8, 9N, 10A, 12F, 15B/C, 22F, 16F, 23A, 31, 18A, 6D, 22A) to serve as a basis for future studies investigating host–pathogen interaction during pneumococcal meningitis. We note that differences in induction of IL-8 between serotypes may be particularly worthy of future study.

Investigate of the Ability of Cronobacter sakazakii Isolated from Clinical Samples of Children Under Two Years to Induce Swimming, Swarming and Biofilm

2018 ◽  
pp. 123
Author(s):  
Luma Abdal Hady Zwein ◽  
Tharieyt Abdulrahman Motlag ◽  
Mohamed Mousa
Keyword(s):  
Cerebrospinal Fluid ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Clinical Samples ◽  
Cronobacter Sakazakii ◽  
Yellow Color ◽  
Golden Yellow ◽  
Vitek 2 ◽  
Biochemical Examination

      The study included 200 samples were collected   from   children  under two   years included (50 samples from each of Cerebrospinal fluid, Blood, Stool and Urine) from, Central Children Hospital and Children's Protections Educational Hospital. Isolates bacterial were obtained cultural, microscopic and biochemical examination and diagnosed to the species by using vitek2 system. The results showed there were contamination in 6.5% of clinical samples. The diagnosed colonies which gave pink color on the MacConkey agar , golden yellow color on the Trypton Soy agar and green color on the Birillent Enterobacter sakazakii agar and gave  a probability of 99% in the vitek 2 and were identified as Cronobacter sakazakii. The identification revealed of thirteen isolates: 6(46.16%) isolated from Cerebrospinal fluid samples, 7(53.84%) isolated from blood samples and not isolated bacteria from stool and urine samples. The results of the investigation of some virulence factors showed that all bacteria isolates were able to swimming with a diameter ranging (1-9 mm) and swarming with a diameter ranging (1-40 mm) and their  ability to biofilm formation  by using three methods. The results show the ability  of  isolates to form biofilm by using  Congo red media  methods where it is 12 (92.30 %) out of 13 isolated bacteria belonging to C. sakazakii  able to form biofilm on the Congo red media  which is 3 (23.07%) were  strong production  biofilm ,   8 (61.53%)  were intermediate  production  biofilm and  1 (7.69% ) were weak  biofilm formation , while the 1 (7.69%)  unable to form biofilm.  Tubes method were all isolates were able to form biofilm, it were found that 3 (23.07%)  isolates strong, and 8 (61.53%) intermediate  and 2( 15.38%)  weak biofilm formation. Microtiter plate method  gave 5 (38.46 %) isolates strong, 6 (46.15%) intermediate and 1 (7.69%) weak biofilm formation.  

scholarly journals A CRISPR-Cas autocatalysis-driven feedback amplification network for supersensitive DNA diagnostics

2021 ◽  
Vol 7 (5) ◽  
pp. eabc7802
Author(s):  
Kai Shi ◽  
Shiyi Xie ◽  
Renyun Tian ◽  
Shuo Wang ◽  
Qin Lu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Genomic Dna ◽  
Reaction Network ◽  
Clinical Samples ◽  
Great Promise ◽  
Base Specificity ◽  
Nucleotide Mutation ◽  
Positive Feedback Circuit ◽  
Stringent Target ◽  
And Function

Artificial nucleic acid circuits with precisely controllable dynamic and function have shown great promise in biosensing, but their utility in molecular diagnostics is still restrained by the inability to process genomic DNA directly and moderate sensitivity. To address this limitation, we present a CRISPR-Cas–powered catalytic nucleic acid circuit, namely, CRISPR-Cas–only amplification network (CONAN), for isothermally amplified detection of genomic DNA. By integrating the stringent target recognition, helicase activity, and trans-cleavage activity of Cas12a, a Cas12a autocatalysis-driven artificial reaction network is programmed to construct a positive feedback circuit with exponential dynamic in CONAN. Consequently, CONAN achieves one-enzyme, one-step, real-time detection of genomic DNA with attomolar sensitivity. Moreover, CONAN increases the intrinsic single-base specificity of Cas12a, and enables the effective detection of hepatitis B virus infection and human bladder cancer–associated single-nucleotide mutation in clinical samples, highlighting its potential as a powerful tool for disease diagnostics.

scholarly journals Comparison of diagnostic clinical samples and environmental sampling for enterovirus and parechovirus surveillance in Scotland, 2010 to 2012

2014 ◽  
Vol 19 (15) ◽  
Author(s):  
H Harvala ◽  
J Calvert ◽  
D Van Nguyen ◽  
L Clasper ◽  
N Gadsby ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Cerebrospinal Fluid ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Environmental Sampling ◽  
Clinical Specimens ◽  
Young Infants ◽  
Coxsackievirus A6 ◽  
The Family ◽  
Common Causes

Human enteroviruses (EV) and parechoviruses (HPeV) within the family Picornaviridae are the most common causes of viral central nervous system (CNS)-associated infections including meningitis and neonatal sepsis-like disease. The frequencies of EV and HPeV types identified in clinical specimens collected in Scotland over an eight-year period were compared to those identified in sewage surveillance established in Edinburgh. Of the 35 different EV types belonging to four EV species (A to D) and the four HPeV types detected in this study, HPeV3 was identified as the most prevalent picornavirus in cerebrospinal fluid samples, followed by species B EV. Interestingly, over half of EV and all HPeV CNS-associated infections were observed in young infants (younger than three months). Detection of species A EV including coxsackievirus A6 and EV71 in clinical samples and sewage indicates that these viruses are already widely circulating in Scotland. Furthermore, species C EV were frequently identified EV in sewage screening but they were not present in any of 606 EV-positive clinical samples studied, indicating their likely lower pathogenicity. Picornavirus surveillance is important not only for monitoring the changing epidemiology of these infections but also for the rapid identification of spread of emerging EV and/or HPeV types.

scholarly journals Quantification of SARS-CoV-2 viral copy number in saliva mouthwash samples using digital droplet PCR

2020 ◽  
Author(s):  
Lisa Oberding ◽  
Jia Hu ◽  
Byron Berenger ◽  
Abu Naser Mohon ◽  
Dylan R. Pillai
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Clinical Samples ◽  
Digital Droplet Pcr ◽  
Viral Copy Number ◽  
Droplet Pcr ◽  
Precise Quantification ◽  
Viral Copy ◽  
Mouth Wash

AbstractSaliva samples were collected through a simple mouth wash procedure and viral load quantified using a technology called digital droplet PCR. Data suggest ddPCR allows for precise quantification of viral load in clinical samples infected with SARS-CoV-2.

A rapid procedure for the isolation of C0t-1 DNA from plants

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 138-142 ◽  
Author(s):  
Michael S. Zwick ◽  
Robert E. Hanson ◽  
M. Nurul Islam-Faridi ◽  
David M. Stelly ◽  
Rod A. Wing ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Copy Number ◽  
Genomic Dna ◽  
Mammalian Species ◽  
Repetitive Dnas ◽  
Rapid Procedure ◽  
Dna Elements ◽  
Repetitive Dna Elements

In situ hybridization (ISH) for the detection of single- or low-copy sequences, particularly large DNA fragments cloned into YAC or BAC vectors, generally requires the suppression or "blocking" of highly-repetitive DNAs. C0t-1 DNA is enriched for repetitive DNA elements, high or moderate in copy number, and can therefore be used more effectively than total genomic DNA to prehybridize and competitively hybridize repetitive elements that would otherwise cause nonspecific hybridization. C0t-1 DNAs from several mammalian species are commercially available, however, none is currently available for plants to the best of our knowledge. We have developed a simple 1-day procedure to generate C0t-1 DNA without the use of specialized equipment.Key words: C0t-1 DNA, in situ hybridization, BACs, plants.

Identification of developmental regulatory genes in Aspergillus nidulans by overexpression.

Genetics ◽  
1995 ◽  
Vol 139 (2) ◽  
pp. 537-547 ◽  
Author(s):  
J F Marhoul ◽  
T H Adams
Keyword(s):  
Growth Inhibition ◽  
Aspergillus Nidulans ◽  
Genomic Dna ◽  
Regulatory Gene ◽  
Regulatory Genes ◽  
Dna Fragments ◽  
Expression Library ◽  
Phenotypic Changes ◽  
Inhibition Of Growth ◽  
Induction Sequence

Abstract Overexpression of several Aspergillus nidulans developmental regulatory genes has been shown to cause growth inhibition and development at inappropriate times. We set out to identify previously unknown developmental regulators by constructing a nutritionally inducible A. nidulans expression library containing small, random genomic DNA fragments inserted next to the alcA promoter [alcA(p)] in an A. nidulans transformation vector. Among 20,000 transformants containing random alcA(p) genomic DNA fusion constructs, we identified 66 distinct mutant strains in which alcA(p) induction resulted in growth inhibition as well as causing other detectable phenotypic changes. These growth inhibited mutants were divided into 52 FIG (Forced expression Inhibition of Growth) and 14 FAB (Forced expression Activation of brlA) mutants based on whether or not alcA(p) induction resulted in accumulation of mRNA for the developmental regulatory gene brlA. In four FAB mutants, alcA(p) induction not only activated brlA expression but also caused hyphae to differentiate into reduced conidiophores that produced viable spores from the tips as is observed after alcA(p)::brlA induction. Sequence analyses of the DNA fragments under alcA(p) control in three of these four sporulating strains showed that in two cases developmental activation resulted from overexpression of previously uncharacterized genes, whereas in the third strain, the alcA(p) was fused to brlA. The potential uses for this strategy in identifying genes whose overexpression results in specific phenotypic changes like developmental induction are discussed.

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