Screening of Infectious Causes of Diarrhea and Genetic Determination of Diarrheagenic E. coli using Multiplex PCR in under 5 Years Children in Egypt

2020 ◽  
Vol EJMM29 (4) ◽  
pp. 27-34
Author(s):  
Yosra M. Hassan ◽  
Saha r M. Khairat ◽  
Nada N. Nawar ◽  
Maha M. Gaafar ◽  
Dina M Hassan ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Infectious Diarrhea ◽  
Genetic Determination ◽  
E Coli ◽  
Single Positive ◽  
Life Threatening ◽  
Stool Specimens ◽  
Genetically Determined ◽  
Microbiological Procedures

Background: Infectious diarrhea represents a life-threatening problem among children in developing countries. Objectives: This work aimed to study bacterial, viral and parasitic causes of acute diarrhea; with genetic determination of diarrheagenic E. coli (DEC) in <5 years children. Methodology: Stool specimens were collected from 206 diarrheal children. Bacterial agents were isolated and identified by standard microbiological procedures. Multiplex PCR was done for genetic determination of DEC subtypes. ELISA was used for detection of viral and parasitic agents. Results: Stool specimens with at least single positive enteropathogen accounted for 98.5% with bacterial, viral and parasitic rates of 98.5%, 42.7% and 25.2%, respectively. Isolated bacteria were DEC (98.5%); Campylobacter (14%), Shigella (3.8%) and Salmonella (1.4%). Rota and Noroviruses showed prevalence of 32.5% and 5.3%, respectively. Conclusion: Infectious diarrhea were mostly due to bacterial agents. DEC and Campylobacter were predominant. EAEC and EPEC were the most genetically determined DEC subtypes.

GENETICS AND METABOLISM

PEDIATRICS ◽  
1960 ◽  
Vol 25 (4) ◽  
pp. 565-571
Author(s):  
Barton Childs
Keyword(s):  
Mutant Gene ◽  
Human Populations ◽  
Affected Patient ◽  
Adrenal Hyperplasia ◽  
Genetic Determination ◽  
The Subject ◽  
Definition Of ◽  
Genetically Determined ◽  
Or Genes

IT IS the part of wisdom when about to give a dissertation which one hopes will at once instruct and entertain, to provide at the outset some definition of the subject matter. Genetics is the study of the heritable components of variation; the heritable factors which determine the range or extent of diversity. So, genetics is concerned with heritable differences and likenesses between individuals and between species. One emphasizes the differences because one can be certain of genetic determination of a particular characteristic only when it exists in a population in two or more alternative forms. It is the variants which catch the eye and hold the attention of the investigator, and which by their presence suggest more than one form of the gene or genes which determine that particular characteristic. I would like in what follows to present some examples of investigations of some aspects of genetics in human populations. ADRENAL HYPERPLASIA Several years ago Dr. Melvin Grumbach and I studied the genetics of adrenal hyperplasia, using as our material the patients of Dr. Lawson Wilkins. Since the disease occurs in more than one member of a sibship and since parents are unaffected, we suspected that it was genetically determined and that the affected patient possessed a double dose of a mutant gene; that is, the characteristic was recessive.

Genetic Determination of Erect and Prostrate Growth Habit in Five Shrubs From Windswept Headlands in the Sydney Region

1992 ◽  
Vol 40 (1) ◽  
pp. 1 ◽  
Author(s):  
TD Auld ◽  
DA Morrison
Keyword(s):  
Growth Habit ◽  
Morphological Characteristics ◽  
Genetic Determination ◽  
Specific Variation ◽  
Banksia Ericifolia ◽  
Growth Habits ◽  
Genetically Determined ◽  
Different Characteristics ◽  
Prostrate Growth

In the Sydney region, many plants from populations on windswept headlands have a more prostrate growth habit compared with plants from populations of the same species occurring away from the coast. To determine whether these different growth habits are genetically determined, plants from four populations of each of five species (Acacia rnyrtifolia, Acacia suaveolens, Banksia ericifolia, Casuarina distyla, Hakea teretifolia) were grown under uniform glasshouse conditions. Multivariate analyses of six morphological characteristics indicate that, for four of these species, the offspring are similar to their maternal parent; we thus conclude that the habit differences are genetically fixed in these populations. The same trend is apparent for <I.C. distyla , although significant variation occurs in the offspring. Univariate analyses indicate that different characteristics reflect the habit differences in different species. For conservation biology, the implications of this intra-specific variation are that attempts should be made to conserve viable populations of all genetically isolated taxa within a species.

scholarly journals Genetic determination of migration strategies in large soaring birds: evidence from hybrid eagles

2018 ◽  
Vol 285 (1884) ◽  
pp. 20180855 ◽  
Author(s):  
Ülo Väli ◽  
Paweł Mirski ◽  
Urmas Sellis ◽  
Mindaugas Dagys ◽  
Grzegorz Maciorowski
Keyword(s):  
Bird Species ◽  
Migration Strategy ◽  
Genetic Determination ◽  
Timing Of Migration ◽  
Habitat Suitability Modelling ◽  
Migration Strategies ◽  
Soaring Birds ◽  
Genetically Determined ◽  
Migratory Movements

The relative contributions of genetic and social factors in shaping the living world are a crucial question in ecology. The annual migration of birds to their wintering grounds and back provides significant knowledge in this field of research. Migratory movements are predominantly genetically determined in passerine birds, while in large soaring birds, it is presumed that social (cultural) factors play the largest role. In this study, we show that genetic factors in soaring birds are more important than previously assumed. We used global positioning system (GPS)-telemetry to compare the autumn journeys and wintering ranges of two closely related large raptorial bird species, the greater spotted eagle Clanga clanga and the lesser spotted eagle Clanga pomarina , and hybrids between them. The timing of migration in hybrids was similar to that of one parental species, but the wintering distributions and home range sizes were similar to those of the other. Tracking data were supported by habitat suitability modelling, based on GPS fixes and ring recoveries. These results suggest a strong genetic influence on migration strategy via a trait-dependent dominance effect, although we cannot rule out the contribution of social interactions.

scholarly journals Detection of 13 Enteric Bacteria and 5 Viruses Causing Acute Infectious Diarrhea Using Multiplex PCR from Direct Stool Specimens

2013 ◽  
Vol 16 (1) ◽  
pp. 33 ◽  
Author(s):  
Seungok Lee ◽  
Yeon-Joon Park ◽  
Hae Kyung Lee ◽  
Soo-Young Kim ◽  
Ja-Young Kim ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Enteric Bacteria ◽  
Infectious Diarrhea ◽  
Stool Specimens

scholarly journals Phenotypic and Genetic Determination of Biofilm Formation in Heat Resistant Escherichia coli Possessing the Locus of Heat Resistance

2021 ◽  
Vol 9 (2) ◽  
pp. 403
Author(s):  
Angela Ma ◽  
Norman Neumann ◽  
Linda Chui
Keyword(s):  
Food Safety ◽  
Heat Resistance ◽  
Food Processing ◽  
Biofilm Formation ◽  
Thermal Inactivation ◽  
Inoculum Size ◽  
Genetic Determination ◽  
E Coli ◽  
Heat Resistant

Despite the effectiveness of thermal inactivation processes, Escherichiacoli biofilms continue to be a persistent source of contamination in food processing environments. E. coli strains possessing the locus of heat resistance are a novel food safety threat and raises the question of whether these strains can also form biofilms. The objectives of this study were to determine biofilm formation in heat resistant E. coli isolates from clinical and environmental origins using an in-house, two-component apparatus and to characterize biofilm formation-associated genes in the isolates using whole genome sequencing. Optimal conditions for biofilm formation in each of the heat resistant isolates were determined by manipulating inoculum size, nutrient concentration, and temperature conditions. Biofilm formation in the heat resistant isolates was detected at temperatures of 24 °C and 37 °C but not at 4 °C. Furthermore, biofilm formation was observed in all environmental isolates but only one clinical isolate despite shared profiles in biofilm formation-associated genes encoded by the isolates from both sources. The circulation of heat resistant E. coli isolates with multi-stress tolerance capabilities in environments related to food processing signify that such strains may be a serious food safety and public health risk.

scholarly journals Application of Multiplex PCR for Detection of Non-O157 Verocytotoxin-Producing Escherichia coli in Bloody Stools: Identification of Serogroups O26 and O111

1998 ◽  
Vol 36 (11) ◽  
pp. 3375-3377 ◽  
Author(s):  
M. Louie ◽  
S. Read ◽  
A. E. Simor ◽  
J. Holland ◽  
L. Louie ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Bloody Stool ◽  
Pcr Assay ◽  
E Coli ◽  
Rapid Assays ◽  
Multiplex Pcr Assay ◽  
Stool Specimens ◽  
Bloody Stools ◽  
Etiologic Diagnosis

Primers were designed to amplify sequences of verocytotoxin genes and eaeA genes of Escherichia coli O26:H11, O111:H8, and O157:H7 in a multiplex PCR assay. This assay successfully detected E. coli O26:H11 in bloody stool specimens in which other enteric pathogens were not detected by culture-based methods. Rapid assays to detect non-O157:H7 verocytotoxin-producingE. coli is important to improve methods for the etiologic diagnosis of hemorrhagic colitis.

Concentration determination of chromatin in unstained resin-embedded sections by means of Z-contrast in STEM

1990 ◽  
Vol 48 (2) ◽  
pp. 186-187
Author(s):  
M. Haider ◽  
B. Bohrmann
Keyword(s):  
Energy Loss ◽  
Freeze Substitution ◽  
Biological Macromolecules ◽  
Local Concentration ◽  
E Coli ◽  
Concentration Determination ◽  
Phosphorous Content ◽  
Z Contrast ◽  
Electron Energy Loss

The technique of Z-contrast in STEM offers the possibility to determine the local concentration of macromolecules like lipids, proteins or DNA. Contrast formation depends on the atomic composition of the particular structure. In the case of DNA, its phosphorous content discriminates it from other biological macromolecules. In our studies, sections of E. coli, the dinoflagellate Amphidinium carterae and Euglena spec. cells were used which were obtained by cryofixation followed by freeze-substitution into acetone with 3% glutaraldehyde. The samples were then embedded either in Lowicryl HM20 at low temperature or in Epon at high temperature. Sections were coated on both sides with 30Å carbon.The DF- and the inelastic image have been recorded simultaneously with a Cryo-STEM. This Cryo-STEM is equipped with a highly dispersive Electron Energy Loss Spectrometer. With this instrument pure Z-contrast can be achieved either with a Filtered DF-image divided by the inelastic image or, as is used in this paper, by dividing the conventional DF-image by an inelastic image which has been recorded with an inelastic detector whose response is dependent on the total energy loss of the inelastically scattered electrons.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

Co-expression of recombinant single chain variable fragment recognizing blood antigen fused with sumo and chaperones in Escherichia coli

2018 ◽  
Vol 40 (4) ◽  
Author(s):  
Dang Thi Ngoc Ha ◽  
Le Thi Thu Hong ◽  
Truong Nam Hai
Keyword(s):  
Recombinant Antibody ◽  
Blood Type ◽  
Soluble Form ◽  
Supernatant Fraction ◽  
Blood Group Antigens ◽  
Single Chain ◽  
E Coli ◽  
Recombinant Scfv ◽  
Single Chain Variable Fragments

Single chain variable fragments (scFv) have widely been used in research, diagnosis and treatment, but the scFv is considered as difficult protein for expression in E. coli. In previous studies, we expressed a construction of recombinant single chain variable fragments again antigen specific for blood type A (antiA-scFv) individually or fused with Trx or SUMO. However, soluble fraction was low abandant and only approximately 40% when fused with Trx, the other cases were expressed in form of inclusion body. Therefore, it was difficult for purification, refolding and activity assesment. In thispaper, we demonstrated a suitable construction for soluble production of antiA-scFv fused with SUMO (SM/antiA-scFv) in presence of chaparones. Under fermentation with 0.1 mM IPTG at 20oC, the SM/antiA-scFv was entirely expressed in soluble form. Importantly, after cleavage from SUMO with SUMOprotease, antiA-scFv was still maintained in the supernatant fraction. Therefore, it can help ensure bioactivity and is useful for purification process. To the best of our knowledge, this is the first report showing soluble recombinant scFv fused with SUMO in presence of chaperone for determination of blood group antigens. Thus, this result facilitates the optimal study of soluble expression, purification and bioactivity determination of the antiA-scFv recombinant antibody. 

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