Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene

1999 ◽  
Vol 84 (1-2) ◽  
pp. 75-83 ◽  
Author(s):  
J.B. Bashiruddin ◽  
C. Cammà ◽  
E. Rebêlo
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Detection ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Babesia Caballi ◽  
Babesia Equi ◽  
Horse Blood ◽  
The 16S Rrna Gene

scholarly journals Biodiversity of Denitrifying and Dinitrogen-Fixing Bacteria in an Acid Forest Soil

2002 ◽  
Vol 68 (8) ◽  
pp. 3818-3829 ◽  
Author(s):  
Christopher Rösch ◽  
Alexander Mergel ◽  
Hermann Bothe
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Nitrite Reductase ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Cloning And Sequencing ◽  
Genetic Traits ◽  
Uncultured Bacteria ◽  
Pcr Products ◽  
The 16S Rrna Gene

ABSTRACT Isolated soil DNA from an oak-hornbeam forest close to Cologne, Germany, was suitable for PCR amplification of gene segments coding for the 16S rRNA and nitrogenase reductase (NifH), nitrous oxide reductase (NosZ), cytochrome cd 1-containing nitrite reductase (NirS), and Cu-containing nitrite reductase (NirK) of denitrification. For each gene segment, diverse PCR products were characterized by cloning and sequencing. None of the 16S rRNA gene sequences was identical to any deposited in the data banks, and therefore each of them belonged to a noncharacterized bacterium. In contrast, the analyzed clones of nifH gave only a few different sequences, which occurred many times, indicating a low level of species richness in the N2-fixing bacterial population in this soil. Identical nifH sequences were also detected in PCR amplification products of DNA of a soil approximately 600 km distant from the Cologne area. Whereas biodiversity was high in the case of nosZ, only a few different sequences were obtained with nirK. With respect to nirS, cloning and sequencing of the PCR products revealed that many false gene segments had been amplified with DNA from soil but not from cultured bacteria. With the 16S rRNA gene data, many sequences of uncultured bacteria belonging to the Acidobacterium phylum and actinomycetes showed up in the PCR products when isolated DNA was used as the template, whereas sequences obtained for nifH and for the denitrification genes were closely related to those of the proteobacteria. Although in such an experimental approach one has to cope with the enormous biodiversity in soils and only a few PCR products can be selected at random, the data suggest that denitrification and N2 fixation are not genetic traits of most of the uncultured bacteria.

scholarly journals Nonspecific PCR Amplification of the 16S rRNA Gene Segment in Different Bacteria by Use of Primers Specific for Campylobacter, Arcobacter, and Helicobacter spp.

2007 ◽  
Vol 45 (4) ◽  
pp. 1376-1377 ◽  
Author(s):  
A. D. Raut ◽  
B. P. Kapadnis ◽  
R. Shashidhar ◽  
J. R. Bandekar ◽  
P. Vaishampayan ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Segment ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
The 16S Rrna Gene

‘Candidatus Liberibacter americanus’, associated with citrus huanglongbing (greening disease) in São Paulo State, Brazil

2005 ◽  
Vol 55 (5) ◽  
pp. 1857-1862 ◽  
Author(s):  
Diva do Carmo Teixeira ◽  
Colette Saillard ◽  
Sandrine Eveillard ◽  
Jean Luc Danet ◽  
Paulo Inácio da Costa ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequence ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Gene Sequences ◽  
16S Rrna Gene Sequences ◽  
Intergenic Regions ◽  
The 16S Rrna Gene

Symptoms of huanglongbing (HLB) were reported in São Paulo State (SPS), Brazil, in March 2004. In Asia, HLB is caused by ‘Candidatus Liberibacter asiaticus' and in Africa by ‘Candidatus Liberibacter africanus’. Detection of the liberibacters is based on PCR amplification of their 16S rRNA gene with specific primers. Leaves with blotchy mottle symptoms characteristic of HLB were sampled in several farms of SPS and tested for the presence of liberibacters. ‘Ca. L. asiaticus' was detected in a small number of samples but most samples gave negative PCR results. Therefore, a new HLB pathogen was suspected. Evidence for an SPS-HLB bacterium in symptomatic leaves was obtained by PCR amplification with universal primers for prokaryotic 16S rRNA gene sequences. The amplified 16S rRNA gene was cloned and sequenced. Sequence analysis and phylogeny studies showed that the 16S rRNA gene possessed the oligonucleotide signatures and the secondary loop structure characteristic of the α-Proteobacteria, including the liberibacters. The 16S rRNA gene sequence phylogenetic tree showed that the SPS-HLB bacterium clustered within the α-Proteobacteria, the liberibacters being its closest relatives. For these reasons, the SPS-HLB bacterium is considered a member of the genus ‘Ca. Liberibacter’. However, while the 16S rRNA gene sequences of ‘Ca. L. asiaticus' and ‘Ca. L. africanus' had 98·4 % similarity, the 16S rRNA gene sequence of the SPS-HLB liberibacter had only 96·0 % similarity with the 16S rRNA gene sequences of ‘Ca. L. asiaticus' or ‘Ca. L. africanus’. This lower similarity was reflected in the phylogenetic tree, where the SPS-HLB liberibacter did not cluster within the ‘Ca. L asiaticus’/‘Ca. L. africanus group’, but as a separate branch. Within the genus ‘Candidatus Liberibacter’ and for a given species, the 16S/23S intergenic region does not vary greatly. The intergenic regions of three strains of ‘Ca. L. asiaticus’, from India, the People's Republic of China and Japan, were found to have identical or almost identical sequences. In contrast, the intergenic regions of the SPS-HLB liberibacter, ‘Ca. L. asiaticus' and ‘Ca. L. africanus' had quite different sequences, with similarity between 66·0 and 79·5 %. These results confirm that the SPS-HLB liberibacter is a novel species for which the name ‘Candidatus Liberibacter americanus' is proposed. Like the African and the Asian liberibacters, the ‘American’ liberibacter is restricted to the sieve tubes of the citrus host. The liberibacter could also be detected by PCR amplification of the 16S rRNA gene in Diaphorina citri, the psyllid vector of ‘Ca. L. asiaticus’, suggesting that this psyllid is also a vector of ‘Ca. L. americanus' in SPS. ‘Ca. L. americanus' was detected in 216 of 218 symptomatic leaf samples from 47 farms in 35 municipalities, while ‘Ca. L. asiaticus' was detected in only 4 of the 218 samples, indicating that ‘Ca. L. americanus' is the major cause of HLB in SPS.

scholarly journals Molecular detection of Anaplasmataceae agents in Dasyprocta azarae in northeastern Brazil

2018 ◽  
Vol 27 (1) ◽  
pp. 98-104 ◽  
Author(s):  
Maria do Socorro Costa de Oliveira Braga ◽  
José Gomes Pereira ◽  
Simone de Jesus Fernandes ◽  
Ingrid Carolinne Lopes Marques ◽  
Renata Passos de Jesus ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Detection ◽  
Evolutionary Model ◽  
Northeastern Brazil ◽  
Likelihood Method ◽  
Rrna Gene ◽  
Nested Pcr Assays ◽  
Tick Vectors ◽  
The 16S Rrna Gene

Abstract Recently, the importance of wild-living rodents for maintenance of pathogens of the family Anaplasmataceae in the environment was investigated. These mammals play a role as reservoirs for these pathogens and act as hosts for the immature stages of tick vectors. The aim of the present study was to investigate the prevalence of Ehrlichia sp. and Anaplasma sp. in 24 specimens of Azara’s agouti (Dasyprocta azarae) that had been trapped in the Itapiracó Environmental Reserve, in São Luís, Maranhão, northeastern Brazil, using molecular methods. Four animals (16.7%) were positive for Ehrlichia spp. in nested PCR assays based on the 16S rRNA gene. In a phylogenetic analysis based on the 16S rRNA gene, using the maximum likelihood method and the GTRGAMMA+I evolutionary model, Ehrlichia sp. genotypes detected in Azara’s agoutis were found to be closely related to E. canis and to genotypes relating to E. canis that had previously been detected in free-living animals in Brazil. The present work showed the first molecular detection of Ehrlichia sp. in Azara’s agoutis in Brazil.

scholarly journals Analysis of 16S rRNA and 51-Kilodalton Antigen Gene and Transmission in Mice of Ehrlichia risticii in Virgulate Trematodes from Elimia livescens Snails in Ohio

2000 ◽  
Vol 38 (9) ◽  
pp. 3349-3358 ◽  
Author(s):  
Manuel Kanter ◽  
Jason Mott ◽  
Norio Ohashi ◽  
Bernard Fried ◽  
Stephen Reed ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Type Strain ◽  
Pcr Analysis ◽  
Composition Analysis ◽  
Rrna Gene ◽  
Antigen Gene ◽  
Ehrlichia Risticii ◽  
Horse Blood ◽  
The 16S Rrna Gene

Operculate snails (the family Pleuroceridae:Elimia livescens) were collected between June and October 1998 from a river in central Ohio where repeated cases of Potomac horse fever (PHF) have occurred. Of collected snails, consistently 50 to 80% carried a combination of cercariae and sporocysts of digenetic virgulate trematodes. The trematodes obtained from each snail were pooled and tested for Ehrlichia risticii, the agent of PHF, by nested PCR using primers specific to the 16S rRNA gene. Out of a total of 209 trematode pools, 50 pools were found to be positive by PCR. The DNA sequence of the 16S rRNA gene identified in one trematode pool was identical to that of the type strain of E. risticii, and the sequence of the gene identified in another pool differed from that of the type strain by 1 nucleotide. Comparison of the deduced amino acid sequence of the partial 51-kDa antigen gene from various sources revealed that Maryland, Ohio (except Ohio 081), and Kentucky strains are in a cluster distinct from the sequences obtained from sources in California and Oregon. Ohio 081 was shown previously by antigenic composition analysis to be distinct from other groups. However, all sequences examined were not segregated according to their sources: horse blood or infected trematodes. E. risticiiwas found to be transmittable from trematodes to mice and was subsequently passaged from infected mice to additional mice, as determined by PCR analysis. Our findings suggest the evolution ofE. risticii in the natural reservoir in separate geographic regions and persistent infection of trematode populations with E. risticii during summer and early fall. The study also suggests that the mouse can be used to isolate E. risticii from the infected trematode.

Detection of Mycoplasma gallinaceum by PCR amplification of the 16S rRNA gene from respiratory disease in village chickens

2018 ◽  
Author(s):  
P. Ponnusamy ◽  
T. Lurthu Reetha ◽  
B.S.M. Ronald ◽  
B. Puvarajan and R. Manicakm
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Respiratory Disease ◽  
Tamil Nadu ◽  
Abdominal Cavity ◽  
Pcr Amplification ◽  
Mycoplasma Species ◽  
Rrna Gene ◽  
Village Chickens ◽  
The 16S Rrna Gene

In the present study, Mycoplasma gallinaceum was detected by PCR amplification of 16S rRNA gene from chronic respiratory disease in village chickens of Cauvery delta region of Tamil Nadu. Necropsy was performed to find out the etiological agent in desi birds mortality. At necropsy, airsacculitis with caseous exudate were found in the thoracic and abdominal cavity. Caseous material from airsacs was collected aseptically from dead birds for detection of Mycoplasma species. DNA was extracted from caseous material by using tissue DNA extraction kit. PCR was carried out using primers to amplify 16S rRNA gene belonging to Mycoplasma species. The amplified product yielded approximately 700-bp length (703 to 713 bp) of the 16S rRNA gene specific for Mycoplasma species. Further, it was subjected to sequence analysis and confirmed as Mycoplasma gallinaceum by NCBI blast analysis. In the present communication, detection of M. gallinaceum by PCR amplification of 16S rRNA gene provides a powerful tool for rapid diagnosis. 

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