scholarly journals Molecular Characterization of Human Rotavirus from Children with Diarrhoeal Disease in Sokoto State, Nigeria

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
B. R. Alkali ◽  
A. I. Daneji ◽  
A. A. Magaji ◽  
L. S. Bilbis ◽  
F. Bande
Keyword(s):  
Monoclonal Antibody ◽  
Diarrhoeal Disease ◽  
Molecular Study ◽  
Rt Pcr ◽  
Human Rotavirus ◽  
Group A ◽  
The Common ◽  
Polymerase Chain ◽  
Pcr Techniques

This study was conducted to detect and characterize prevalent human group A rotavirus strains from 200 diarrheic children in Sokoto, Nigeria, by ELISA, monoclonal antibody (Mab) serotyping and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) techniques. Rotavirus was detected in 25.5% of the children. The G-serotypes observed in circulation were G4: 16 (59.3%), G1: 4 (14.8%), G2: 3 (11.1%), G3: 3 (11.1%), and G12: 1 (3.7%). The monoclonal antibody (Mab) serotyping detected G1 and G3 but did not detect G4 and G2 serotypes. The Mab typing of the G1 and G3 serotypes was consistent with the result of the RT-PCR. The VP4 genotypes detected were P[6] 3 (13%), P[8] 11 (47.8%), and the rare human P genotype (P[9]), found in 9 patients (39.1%). Nine strains identified with the common G and P combinations were G4 P[8] 5 (56%), G4 P[6] 1 (11%), G1 P[8] 2 (22%), and G3 P[8] 1 (11%), while seven strains with unusual combinations or rare G or P genotypes identified were G12 P[8] 1 (14%), G2 P[8] 2 (29%), and G4 P[9] 4 (57%). To our knowledge this is the first molecular study of human rotavirus and report of rare human G and P serotypes in Sokoto State.

scholarly journals Molecular characterization of a rare G1P[19] rotavirus strain from India: evidence of reassortment between human and porcine rotavirus strains

2009 ◽  
Vol 58 (12) ◽  
pp. 1611-1615 ◽  
Author(s):  
Shobha D. Chitambar ◽  
Ritu Arora ◽  
Preeti Chhabra
Keyword(s):  
Amino Acid Sequences ◽  
Rt Pcr ◽  
Coding Regions ◽  
Subgroup Ii ◽  
Human Rotavirus ◽  
Independent Segregation ◽  
Group A ◽  
Porcine Rotavirus ◽  
Porcine Origin

This study pertains to the characterization of a human rotavirus strain (NIV929893) with a rare specificity of G1P[19]. Three structural genes (VP4, VP6 and VP7) and one non-structural gene (NSP4) of strain NIV929893 were subjected to RT-PCR for amplification of entire coding regions. All of the amplicons were sequenced to carry out phylogenetic analysis. The complete amino acid sequences of the VP7 and VP4 gene products showed clustering of the VP7 gene with G1 strains of human origin and the VP4 gene with P[19] strains of porcine origin. The two viral proteins VP6 and NSP4, described previously as genetically linked proteins, were shown to be subgroup II and genotype B of human and porcine origins, respectively. The findings of this study provide evidence of reassortment between VP7/VP6 genes of humans and VP4/NSP4 genes of porcine species and an independent segregation of VP6 and NSP4 genes in a group A human rotavirus strain with G1P[19] specificity.

scholarly journals Molecular Characterization of VP6 Genes of Human Rotavirus Isolates: Correlation of Genogroups with Subgroups and Evidence of Independent Segregation

2002 ◽  
Vol 76 (13) ◽  
pp. 6596-6601 ◽  
Author(s):  
Miren Iturriza Gómara ◽  
Cecilia Wong ◽  
Sandra Blome ◽  
Ulrich Desselberger ◽  
Jim Gray
Keyword(s):  
Molecular Characterization ◽  
Rt Pcr ◽  
Genetic Lineages ◽  
Reverse Transcription Pcr ◽  
Human Rotavirus ◽  
Independent Segregation ◽  
Genes Encoding ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT A reverse transcription-PCR (RT-PCR) was established to amplify a 379-bp cDNA fragment (nucleotides 747 to 1126, coding for amino acids 241 to 367) of the VP6 gene of group A rotaviruses associated with subgroup (SG) specificity. Thirty-eight human rotavirus strains characterized with SG-specific monoclonal antibodies were subjected to VP6-specific RT-PCR, and PCR amplicons were used for sequencing. Nucleic acid sequencing and phylogenetic analysis of the VP6 amplicons revealed two clusters, or genogroups. Two genetic lineages were distinguished within genogroup I, consisting of strains serologically characterized as SG I, and three genetic lineages were distinguished within genogroup II, composed of strains serologically characterized as SG II, SG I + II, and SG non-I, non-II. Subgrouping of rotaviruses by means of serological methods may result in strains not being assigned the correct SG or in a failure of strains to subgroup. Molecular characterization of the SG-defining region of VP6 provided evidence for independent segregation of the rotavirus genes encoding VP4, VP6, and VP7.

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Immunohistochemical Analysis ◽  
Specific Protein ◽  
Platelet Membrane ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Megakaryocytic Cell

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

scholarly journals Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

2012 ◽  
Vol 3 (1) ◽  
pp. 13
Author(s):  
Aline T.A. Chagas ◽  
Michelle D. Oliveira ◽  
Jose M.S. Mezencio ◽  
Eduardo A.M. Silva ◽  
Leandro L. Oliveira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Epidemiological Surveillance ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Denv Serotypes ◽  
Polymerase Chain ◽  
Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

Characterization of the NSP4 gene of group A human rotavirus G1P[8] strains circulating in Sapporo, Japan from 1987 to 2000

2013 ◽  
Vol 86 (2) ◽  
pp. 354-359 ◽  
Author(s):  
Masatoshi Tatsumi ◽  
Yoshinobu Nagaoka ◽  
Takeshi Tsugawa ◽  
Yuko Yoto ◽  
Tsukasa Hori ◽  
...  
Keyword(s):  
Nsp4 Gene ◽  
Human Rotavirus ◽  
Group A

scholarly journals Molecular characterization of group A rotaviruses detected in children with gastroenteritis in Ireland in 2006–2009

2011 ◽  
Vol 140 (2) ◽  
pp. 247-259 ◽  
Author(s):  
O. CASHMAN ◽  
P. J. COLLINS ◽  
G. LENNON ◽  
B. CRYAN ◽  
V. MARTELLA ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Irish Population ◽  
Surveillance Activity ◽  
Human Rotavirus ◽  
Group A Rotaviruses ◽  
Group A ◽  
Hospital Acquired ◽  
Rotavirus Infections ◽  
Pcr Genotyping

SUMMARYCommunity and hospital-acquired cases of human rotavirus are responsible for millions of gastroenteritis cases in children worldwide, chiefly in developing countries, and vaccines are now available. During surveillance activity for human rotavirus infections in Ireland, between 2006 and 2009, a total of 420 rotavirus strains were collected and analysed. Upon either PCR genotyping and sequence analysis, a variety of VP7 (G1–G4 and G9) and VP4 (P[4], P[6], P[8] and P[9]) genotypes were detected. Strains G1P[8] were found to be predominant throughout the period 2006–2008, with slight fluctuations seen in the very limited samples available in 2008–2009. Upon either PCR genotyping and sequence analysis of selected strains, the G1, G3 and G9 viruses were found to contain E1 (Wa-like) NSP4 and I1 VP6 genotypes, while the analysed G2 strains possessed E2 NSP4 and I2 VP6 genotypes, a genetic make-up which is highly conserved in the major human rotavirus genogroups Wa- and Kun-like, respectively. Upon sequence analysis of the most common VP4 genotype, P[8], at least two distinct lineages were identified, both unrelated to P[8] Irish rotaviruses circulating in previous years, and more closely related to recent European humans rotaviruses. Moreover, sequence analysis of the VP7 of G1 rotaviruses revealed the onset of a G1 variant, previously unseen in the Irish population.

Clinical presentation and molecular characterization of group B rotaviruses in diarrhoea patients in Bangladesh

2011 ◽  
Vol 60 (4) ◽  
pp. 529-536 ◽  
Author(s):  
Farjana Saiada ◽  
H. N. Ashiqur Rahman ◽  
Sayra Moni ◽  
M. Manjurul Karim ◽  
Mahmoud Reza Pourkarim ◽  
...  
Keyword(s):  
Continuous Monitoring ◽  
Diarrhoeal Disease ◽  
Specific Pattern ◽  
Severe Dehydration ◽  
Animal Group ◽  
Group A ◽  
Stool Samples ◽  
Group B ◽  
Rotavirus Infections

A total of 1106 stool samples collected from diarrhoea patients admitted to Dhaka hospital of the International Centre for Diarrhoeal Disease Research, Bangladesh, during January–December 2008 were analysed for the presence of rotavirus-specific RNA by PAGE. The group B-specific RNA migration pattern was detected in 26 patients (2.4 %) and group A-specific pattern in 259 patients (23.4 %). Clinical data from group A and group B rotavirus-infected patients indicated that episodes did not differ much in the prevalence of diarrhoea, number of stools, outcome or differences in gender. However, abdominal pain was more common in group B rotavirus infections (36 vs 15 %, P=0.02) and the virus was responsible for more severe dehydration compared with group A-infected patients (12 vs 3 %, P=0.04). Sequence analyses of VP4, VP7 and NSP2 indicated that an Indian–Bangladeshi lineage of the virus, which is different from both the prototype (Chinese) lineage and from the animal group B rotaviruses, has been circulating in Bangladesh. Continuous monitoring of group B rotaviruses both in hospitals and in the community will be helpful to determine the true burden of group B rotaviruses.

Molecular characterization of VP6-encoding gene of group A human rotavirus samples from central west region of Brazil

2008 ◽  
Vol 80 (11) ◽  
pp. 2034-2039 ◽  
Author(s):  
Talissa de Moraes Tavares ◽  
Wilia Marta Elsner Diederichsen de Brito ◽  
Fabíola Souza Fiaccadori ◽  
Juliana Alves Parente ◽  
Paulo Sérgio Sucasas da Costa ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
West Region ◽  
Human Rotavirus ◽  
Group A ◽  
Encoding Gene

Isolation and Characterization of Mesenchymal Stem Cells from Chicken Liver

2020 ◽  
Vol 10 (1) ◽  
pp. 8-16
Author(s):  
Xibin Liu ◽  
Shuang Zhang ◽  
Weijun Guan ◽  
Dong Zheng
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Protein Markers ◽  
Multilineage Differentiation ◽  
Rt Pcr ◽  
Multipotent Stem Cells ◽  
Isolation And Characterization ◽  
Potential Applications ◽  
Polymerase Chain

Hepatic mesenchymal stem cells (HMSCs) are multipotent stem cells that is a vital part of the regeneration of hepatocytes after injury. In this study, HMSCs were isolated in embryonic livers from of 12-day-old chick embryo using collagenase, and the primary HMSCs were sub-cultured to passage. The protein markers of HMSCs, namely CD71, CD29 and CD44, were tested with immunofluorescence and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The proliferation of HMSCs in different passages was detected using growth curve, which shown a typically sigmoidal. And then, the pluripotent of HMSCs was analyzed, the results showed that HMSCs could directly induce to differentiate into neural-like cells, adipocytes, and osteoblasts. Our data illustrated that the chick HMSCs have same characteristics to those obtained from other species. The capacity of these cells for multilineage differentiation shows promise for many potential applications.

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