Identification Of New Genotypes Research Articles

Detection and Molecular Characterization of Porcine Teschoviruses in India: Identification of New Genotypes

Detection and Molecular Characterization of Porcine Teschoviruses in India: Identification of New Genotypes

Identification of Modern High-Yield Soybean Genotypes for Potassium-Use Efficiency in Sandy Soil of the Brazilian Cerrado

Soybean is the main leguminous crop in Brazil, mostly grown in tropical soils with low potassium (K) availability. Therefore, the identification of new genotypes with efficient K uptake and utilization in environments with low exchangeable K content is an economically viable alternative to maximize crop yield in Brazil. A study was conducted to investigate the response of 25 modern high-yield soybean genotypes for K-use efficiency in a sandy tropical soil of the Brazilian Cerrado. Treatments were distributed in a completely randomized design in a 2 × 25 factorial scheme: two levels of K fertilization [20 mg K dm−3 (low level) or 200 mg K dm−3 (high level)] and 25 soybean genotypes with three replicates. Plant morphological traits, leaf K, and crop production components were measured. Based on grain production data, K-use efficiency (KUE) and response efficiency (RE) to K fertilization were calculated. Leaf area, shoot dry matter, pod number per plant, 1000-grain weight, and grain yield were the crop characteristics most limited by low soil K availability. The soybean genotypes “TMG7061 IPRO”, “BMX Bônus IPRO”, “RK6719 IPRO”, and “RK8317 IPRO” were classified as efficient in the use of soil K and are the most suitable genotypes to be cultivated in agricultural soils with low K availability. The genotypes “98R35 IPRO”, “HO Maracaí IPRO”, “BMX Bônus IPRO”, and “RK7518 IPRO” were classified as responsive to K fertilization and are the most recommended genotypes for cultivation in agricultural areas with the application of high K fertilizer rates. The genotype “BMX Bônus IPRO” simultaneously combines characteristics of K-use efficiency and response to K fertilization and hence can be grown in both K-deficient and optimal soils.

Enterocytozoon bieneusi genotypes in cats and dogs in Victoria, Australia

BackgroundEnterocytozoon bieneusi is one of the commonest microsporidians contributing to human microsporidiosis, and is frequently found in animals in various countries. However, there is limited epidemiological information on this microorganism in Australia. Here, we undertook the first molecular epidemiological study of E. bieneusi in cats and dogs in Victoria.ResultsGenomic DNAs were extracted from 514 individual faecal deposits from cats (n = 172) and dogs (n = 342) and then tested using PCR-based sequencing of the internal transcribed spacer (ITS) of nuclear ribosomal DNA. Four distinct genotypes (designated D, PtEb IX, VIC_cat1 and VIC_dog1) of E. bieneusi were identified in 20 of the 514 faecal samples (3.9%). Genotype D is known to have a broad host range (humans and other animals) and has a wide geographical distribution around the world. The identification of this genotype here suggests that companion animals might represent reservoir hosts that are able to transmit E. bieneusi infection to humans in Australia. A phylogenetic analysis of ITS sequence data revealed that the novel genotype VIC_cat1 is related to the known genotype type IV within Group 1, and the new genotype VIC_dog1 is linked to a contentious “Group 3”, which includes genotypes reported previously in the published literature to represent Group 2 or 3.ConclusionsA future, large-scale phylogenetic study of all known E. bieneusi genotypes, including VIC_dog1, should aid in clarifying their relationships and assignment to Groups, and in the identification of new genotypes, thus assisting epidemiological investigations.

Genetic variability of the measles virus hemagglutinin gene in B3 genotype strains circulating in Northern Italy

Sequencing the whole measles virus hemagglutinin (H) gene, in conjunction with a 450-nucleotide region of the nucleoprotein gene (N-450), is helpful for the identification of new genotypes and as an auxiliary in outbreak characterization. In addition, it is essential to be able to predict the antigenic changes of the H protein to gain a better monitoring of the response to the vaccine. In this study, we obtained the full-length H gene sequences from 19 measles virus (MV) strains belonging to two B3 genotype variants circulating in Lombardy (Northern Italy) between July 2015 and February 2016 and evaluated the variability of the whole MV-H gene. Furthermore, we compared the obtained H amino acid sequences to all MV sequences available in the GenBank database (n = 1152 in total) and analyzed the amino acid substitutions in the H protein within clades where the Italian strains were included. We identified a higher variability in the H gene compared to the N-450 region and our results support previous studies, highlighting that the H gene is more informative for characterizing the MV B3 genotype than the N-450 sequence. Some of the amino acid substitutions were fixed in the viral population and, remarkably, some of the amino acid substitutions were typically present only in the Italian sequences. Accumulating further molecular information about MV-H gene will be necessary to enable in-depth analyses of the variability of this gene in the vaccinated population.

Molecular characterization of echovirus 13 uncovering high genetic diversity and identification of new genotypes in Pakistan

Echovirus 13 (E-13) is reported worldwide and is mostly related to aseptic meningitis but it is also isolated from cases of acute flaccid paralysis (AFP). Unfortunately, all studies conducted on non polio enterovirus in Pakistan only confirm E-13 isolation based on microneutralization assay but there is lack of molecular epidemiological data on this serotype. In this study, 113 stool samples were collected from AFP patients during 2008–2010. An enterovirus primer mediated real-time reverse transcriptase polymerase chain reaction, a standard microneutralization assay and sequencing of viral protein 1 gene (VP1) identified the predominant serotype E-13. For molecular characterization, genetic relationship between 12 clinical isolates of echovirus 13 was investigated by partial sequencing of viral protein 1 gene. These strains, combined with related sequences from GenBank were divided phylogenetically into two different genogroups A and B (>30% divergence) and were found genetically distinct from the circulating strains in the world. Additionally, phylogenic grouping pattern revealed that the study strains clustered into three distinct subgroups (A3, A7 and B3) having >23% nucleotide divergence representing three new genotypes. The genotype A7 seems to be restricted geographically. In conclusion, the current study provides an overview of the molecular epidemiology and evolution of E-13 in the country. This study strongly suggests that enterovirus surveillance system should be established in the country to determine the temporal and geographical trends and disease pattern of different enterovirus serotypes in the community.

Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran.

The continuous increase in Acanthamoeba keratitis, a severe corneal infection, worldwide is mainly due to the increase in the number of soft contact lens users. In the present study, which involves a 5-year study, a total of 138 corneal scraps and contact lenses together with their paraphernalias were obtained from suspected amoebic keratitis patients. All samples were cultured using culture-enrichment method. Pathogenic assay, using thermotolerance and osmotolerance tests were also performed on the positive strains. Sequencing of the isolated strains was done by targeting the DF3 region of 18s rRNA gene. The results revealed that 18 (13%) of patients were infected with Acanthamoeba spp. As expected, T4 genotype was the most common genotype among the clinical samples; however, in three cases, Acanthamoeba belonging to T11 and T9 were detected. Interestingly, T9 genotype, commonly classified as non-pathogenic amoebae, was identified as a causal agent of a patient with amoebic keratitis. From the pathogenic assay, four strains belonging to T4 genotypes were highly pathogenic. This is the first report of Acanthamoeba T9 genotypes isolated in Iran and the first report of T9 type occurring in amoebic keratitis patients worldwide. Due to the increasing trend of amoebic keratitis (AK) and the identification of new genotypes, such as T9 as the causative agent of AK, more researches in this field are necessary in the region and the world at large.

Identification of new genotype of Echovirus 19 from children with Acute Flaccid Paralysis in Pakistan.

Enteroviruses are known to cause childhood paralysis. The purpose of this study was to examine the genetic diversity and to determine the association of non-polio enteroviruses (NPEVs) with acute flaccid Paralysis (AFP). Stool samples (n = 1191) of children with AFP were collected from Khyber Pakhtunkhwa and Federally Administered Tribal Areas of Pakistan. Poliovirus was isolated in 205 (17.2%) samples and NPEVs were found in 215 (18.0%) samples. Out of 215 viruses, 124 (57.7%) were typed into 19 different types of enteroviruses while 91 (42.3%) remained untypeable on microneutralization assay that were reconfirmed as NPEVs by real time PCR. Echovirus 19 (20/35; 57.1%) was found the most prevalent type based on VP1 nucleotide sequencing with increased genetic diversity. Phylogenetic analysis revealed the circulation of a new genotype of E-19 in the country. The findings of this study are of great importance for future research and propose to establish the enterovirus surveillance system in the country to readily identify more enteroviruses and to monitor the emergence of new variants/genotypes especially at the moment when we are at the verge of polio eradication phase.

Detection of Acinetobacter baumannii in human head and body lice from Ethiopia and identification of new genotypes

Acinetobacter baumannii has previously been detected and genotyped in human body lice. The objectives of this study were to determine the presence of this bacterium in head and body lice collected from healthy individuals in Ethiopia by molecular methods and to characterize the genotype. Human lice from locations at different altitudes in Ethiopia were screened for the presence of Acinetobacter sp by targeting the rpoB gene. Acinetobacter baumannii was detected and genotyped using recA PCR amplification. A total of 115 head and 109 body lice were collected from 134 healthy individuals. Acinetobacter sp were found in 54 head (47%) and 77 body (71%) lice. The recA gene was sequenced for 60 of the Acinetobacter sp and 67% were positive for A. baumannii; genotype 1 was retrieved the most frequently. Our study is the first to show the presence of A. baumannii in human body lice, and also in head lice, in Ethiopia.

Global Analysis of Sequence Diversity within HIV-1 Subtypes across Geographic Regions

AIMS: HIV-1 sequence diversity can affect host immune responses and phenotypic characteristics such as antiretroviral drug resistance. Current HIV-1 sequence diversity classification uses phylogeny-based methods to identify subtypes and recombinants, which may overlook distinct subpopulations within subtypes. While local epidemic studies have characterized sequence-level clustering within subtypes using phylogeny, identification of new genotype - phenotype associations are based on mutational correlations at individual sequence positions. We perform a systematic, global analysis of position-specific pol gene sequence variation across geographic regions within HIV-1 subtypes to characterize subpopulation differences that may be missed by standard subtyping methods and sequence-level phylogenetic clustering analyses. MATERIALS #ENTITYSTARTX00026; METHODS: Analysis was performed on a large, globally diverse, cross-sectional pol sequence dataset. Sequences were partitioned into subtypes and geographic subpopulations within subtypes. For each subtype, we identified positions that varied according to geography using VESPA (viral epidemiology signature pattern analysis) to identify sequence signature differences and a likelihood ratio test adjusted for multiple comparisons to characterize differences in amino acid (AA) frequencies, including minority mutations. Synonymous nonsynonymous analysis program (SNAP) was used to explore the role of evolutionary selection witihin subtype C. RESULTS: In 7693 protease (PR) and reverse transcriptase (RT) sequences from untreated patients in multiple geographic regions, 11 PR and 11 RT positions exhibited sequence signature differences within subtypes. Thirty six PR and 80 RT positions exhibited within-subtype geography-dependent differences in AA distributions, including minority mutations, at both conserved and variable loci. Among subtype C samples from India and South Africa, nine PR and nine RT positions had significantly different AA distributions, including one PR and five RT positions that differed in consensus AA between regions. A selection analysis of subtype C using SNAP demonstrated that estimated rates of nonsynonymous and synonymous mutations are consistent with the possibility of positive selection across geographic subpopulations within subtypes. CONCLUSION: We characterized systematic genotypic pol differences across geographic regions within subtypes that are not captured by the subtyping nomenclature. Awareness of such differences may improve the interpretation of future studies determining the phenotypic consequences of genetic backgrounds.

Geographical and genetic diversity of the human hepatitis B virus

Hepatitis B virus (HBV) is one of the most widely distributed viruses that infect humankind. Distinct clinical and virological characteristics of the HBV-infection have been reported in different geographical parts of the world and are increasingly associated with genetic diversity of the infecting virus. HBV is classified into genotypes and subgenotypes that are associated with ethnicity and geography. The genetic diversity of HBV in its various aspects has been the subject of extensive investigations during the last few decades. Since molecular epidemiology research tools have become widely available, the number of new publications in this field has grown exponentially. This review summarises the recent publications on the geographical distribution of genetic variants of HBV, and proposes updated criteria for the identification of new genotypes and subgenotypes of the virus.

A statistical model for unwarping of 1-D electrophoresis gels

A statistical model is proposed which relates density profiles in 1-D electrophoresis gels, such as those produced by pulsed-field gel electrophoresis (PFGE), to databases of profiles of known genotypes. The warp in each gel lane is described by a trend that is linear in its parameters plus a first-order autoregressive process, and density differences are modelled by a mixture of two normal distributions. Maximum likelihood estimates are computed efficiently by a recursive algorithm that alternates between dynamic time warping to align individual lanes and generalised-least-squares regression to ensure that the warp is smooth between lanes. The method, illustrated using PFGE of Escherichia coli O157 strains, automatically unwarps and classifies gel lanes, and facilitates manual identification of new genotypes.

Genotyping HCV isolates from Italy by type-specific PCR assay in the core region

A revision of the polymerase chain reaction (PCR) core procedure was performed for genotyping hepatitis C virus (HCV) in 139 patients from Italy. This procedure, developed prior to the identification of new genotypes, may be inadequate in several geographical areas. We proposed a new typing mixture in which primers for types 2c and 4, that are reported to be circulating in Italy, were added and a primer for type 2b was substituted. Using the modified procedure, 139 HCV-positive patients were analysed. The HCV genotype was identified in 96.4% of the cases. We observed double infections and unclassified genotypes in 5 (3.6%) and 5 (3.6%) patients, respectively. The classification of isolates into genotypes and subtypes 2b, 2c and 4 was confirmed by sequence analysis. Furthermore, the efficiency and accuracy of the modified core procedure were evaluated by parallel testing of 107 out of 139 samples using the line probe assay, and demonstrated a 98.9% degree of concordance. The results demonstrated the specificity of the selected primers for type 2c, 2b and 4 and confirmed the circulation of types 2c and 4 in Italy. In conclusion, the proposed modified PCR procedure is the only primer-specific PCR genotyping method available for identification of the 2c and 4 genotypes reported to be circulated in Italy and other European countries.