scholarly journals Detection and molecular characterisation of Ehrlichia canis in naturally infected dogs in South West Nigeria

2018 ◽  
Vol 66 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Olukayode Olugbenga Daramola ◽  
Michael Irewole Takeet ◽  
Ibironke Kofoworola Oyewusi ◽  
Mufutau Atanda Oyekunle ◽  
Adewale Oladele Talabi
Keyword(s):  
Ehrlichia Canis ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Rickettsial Disease ◽  
Blood Samples ◽  
Canine Ehrlichiosis ◽  
South West ◽  
Polymerase Chain ◽  
Reference Sequences ◽  
The 16S Rrna Gene

Canine ehrlichiosis is an important tick-borne rickettsial disease mainly caused by Ehrlichia canis. This study aimed to detect and characterise E. canis in dogs in Abeokuta, Nigeria by microscopy and nested PCR. Blood samples were collected from 205 dogs, thin smears were made, field-stained, and DNA was extracted from the blood samples. A partial region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced unidirectionally. Ehrlichial morulae were detected in three dogs (1.5%). The PCR test revealed that 47 dogs (22.9%) were positive for E. canis. The lengths of the sequences obtained range from 374 bp to 376 bp with an average G-C content of 37% and 98–99% homology with the reference sequences in GenBank. The aligned autochthonous sequences were less polymorphic. The phylogenetic analysis separated sequences reported previously in Nigeria from the autochthonous sequences. The present work shows that the strain of E. canis detected in the study area is genetically different from those reported in the northern part of Nigeria and more closely related to sequences from Brazil and India.

scholarly journals Detection and genetic characterization of "Candidatus Mycoplasma haemomacaque" infection among long-tailed macaques (Macaca fascicularis) in Thailand using broad-range nested polymerase chain reaction assay

2021 ◽  
Vol 14 (4) ◽  
pp. 943-948
Author(s):  
Wanat Sricharern ◽  
Supakarn Kaewchot ◽  
Sarawan Kaewmongkol ◽  
Natnaree Inthong ◽  
Thitichai Jarudecha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Genetic Characterization ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

Background and Aim: Hemoplasmas are defined as small, epicellular parasitic bacteria that can infect the red blood cells of several mammalian species. Diseases caused by these bacteria range from asymptomatic infections to acute hemolytic anemia. However, data on hemoplasmas in non-human primates in Thailand remain to be limited. Therefore, this study aims to determine the occurrence and genetic diversity of hemoplasmas among long-tailed macaques in Thailand. Materials and Methods: Blood samples were collected from 339 long-tailed macaques in three provinces of Thailand. DNA was then extracted from the blood samples and tested for hemoplasma using broad-range nested polymerase chain reaction (PCR) based on the 16S rRNA gene. PCR-positive samples were sequenced, and phylogenetic analysis for species identification was conducted. Results: In total, 38 (11.2%) out of the 339 samples were found to be positive for hemoplasmas, based on the broad-range nested PCR assay of the 16S rRNA gene. The 16S rRNA sequences of Mycoplasma spp. were highly similar (98-99% identity) to "Candidatus Mycoplasma haemomacaque." Furthermore, phylogenetic analysis using maximum likelihood demonstrated that the sequences were located in the same cluster of "Ca. M. haemomacaque." Conclusion: The detection of hemoplasmas among long-tailed macaques in Thailand is reported. Genetic characterization confirmed that these hemoplasmas are closely related to "Ca. M. haemomacaque." These results indicate that long-tailed macaques in several locations in Thailand may be infected and serve as reservoirs for this parasite.

scholarly journals Identification of Mycoplasma genitalium from clinical swabs by direct PCR

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1993
Author(s):  
Robinson M. Irekwa ◽  
Perpetual Ndung'u ◽  
Peter Kipkemboi ◽  
Tonny Teya ◽  
Anne Wanjiru Mwangi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Workers ◽  
Genital Tract ◽  
Mycoplasma Genitalium ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Direct Pcr ◽  
The World ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

Mycoplasma genitalium is one of the smallest self-replicating organisms. It is an obligate parasite found in the human genital tract. In men, the bacteria cause both acute and chronic non-gonococcal urethritis (NGU). In women, it has been associated with pelvic inflammatory disease and cervicitis among other related infections. Treatment of M. genitalium related infections has been effective using antibiotics such as the macrolides (e.g. azithromycin) and fluoroquinolones. However, there have been recorded cases of resistance to these antibiotics in various parts of the world as a result of a mutation in the 23SrRNA gene, although the antibiotic resistance has not been well established. The aim of this study was to detect M. genitalium in 352 swab samples collected from a clinic for sex workers in Nairobi, Kenya. DNA was extracted from the swabs and stored as a crude extract at -31°C. The swab lysates were subjected to direct polymerase chain reaction using primers that specifically target the 16S rRNA gene for M. genitalium. A total of 29 samples tested positive for M. genitalium. The data results showed a M. genitalium prevalence of 8.24% among sex workers in Nairobi, Kenya.

scholarly journals Molecular and serological detection of Ehrlichia canis in naturally exposed dogs in Iran: an analysis on associated risk factors

2014 ◽  
Vol 23 (1) ◽  
pp. 16-22 ◽  
Author(s):  
Nadi Maazi ◽  
Abdolali Malmasi ◽  
Parviz Shayan ◽  
Seyed Mahdi Nassiri ◽  
Taghi Zahraei Salehi ◽  
...  
Keyword(s):  
Risk Factors ◽  
Rural Areas ◽  
Ehrlichia Canis ◽  
Rrna Gene ◽  
Igg Antibodies ◽  
Serological Detection ◽  
Blood Samples ◽  
Associated Risk Factors ◽  
First Time ◽  
The 16S Rrna Gene

The general aim of this study, which was conducted for the first time in Iran, was to evaluate the seroprevalence and geographical distribution of Ehrlichia canis in a dog population in Iran, followed by molecular confirmation using PCR and sequencing. Blood samples were collected from 240 dogs in different areas of Alborz and Tehran Provinces and initially analyzed using the immunofluorescent antibody (IFA) test to detect anti-Ehrlichia canis IgG antibodies. Subsequently, nested PCR was performed based on a fragment of the 16S rRNA gene of E. canis on serologically positive samples. The results showed that 40/240 dogs (16.6%) presented anti-Ehrlichia canis IgG antibodies and that nine of the blood samples from the 40 seropositive dogs (22.5%) contained E. canis DNA, which was confirmed by sequencing. The seroprevalence of E. canis tended to be higher in purebred, one to three-year-old male dogs living in the Plain zone, in rural areas; however, this difference was not statistically significant.

scholarly journals Detection of Genetically Modified Corn (Bt176) in Spiked Cow Blood Samples by Polymerase Chain Reaction and Immunoassay Methods

2005 ◽  
Vol 88 (2) ◽  
pp. 654-664 ◽  
Author(s):  
Laetitia Petit ◽  
Fabienne Baraige ◽  
Yves Bertheau ◽  
Philippe Brunschwig ◽  
Annick Diolez ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Cry1ab Protein ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

Abstract The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction–enzyme-linked immunosorbent assay (PCR–ELISA) and the 5′-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (αs1-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.

scholarly journals Molecular detection of Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania

2020 ◽  
Author(s):  
Noel Gahamanyi ◽  
Leonard E.G. Mboera ◽  
Mecky I. Matee ◽  
Dieudonné Mutangana ◽  
Raghavendra G. Amachawadi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Faecal Samples ◽  
Rrna Sequencing ◽  
Polymerase Chain ◽  
Campylobacter Species ◽  
The 16S Rrna Gene

Abstract Background: A growing number of Campylobacter species other than C. jejuni and C. coli have been considered as emerging human and animal pathogens. However, the contribution of these species to human gastroenteritis is poorly documented. This study aimed at detecting Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania using Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, and Sanger sequencing . Methods: A total number of 100 faecal samples (70 from human and 30 from cattle) were collected from diarrheic and non-diarrheic patients and healthy cattle in Kilosa district, Tanzania from July to October 2019. Species identification was conducted by PCR and 16S rRNA sequencing. The phylogenetic analysis was carried out by comparison of the 16S rRNA gene sequences to reference strains by the Neighbor-Joining method in MEGA X. Results: Campylobacter species detection rate by PCR was 65.7% (46/70) and 20% (6/30) in humans and cattle, respectively. There were five human diarrheic cases, four showed Campylobacter presence and two were from children ≤15 years of age. In humans, the 16S rRNA sequencing revealed that C. concisus was the most predominant species occurring at a frequency of 37.8% (14/37), followed by uncultured Campylobacter spp. 24.3% (9/37) and C. hominis 21.6% (8/37). The least represented species were C. jejuni and C. lanienae all occurring at 2.7% (1/37). In cattle, five (100%) sequenced PCR products matched with C. lanienae . Phylogenetic analysis revealed that Campylobacter 16S rRNA sequences were closely related to C. concisus , uncultured Campylobacter sp., C. hominis , and C. gracilis . Conclusion: The non- C. jejuni / C. coli species are present in human and cattle faecal samples and their true occurrence is probably under-reported due to shortcomings of conventional techniques used in most diagnostic microbiology laboratories. Based on our findings, we recommend that molecular techniques be adopted for direct detection of Campylobacter species during routine laboratory screening and surveillance studies. Keywords: Campylobacter , molecular diagnostics, polymerase chain reaction, sequencing, gastroenteritis, Tanzania

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