scholarly journals Genotype diversity of Mycoplasma Hyopneumoniae in Chinese swine herds based on multilocus sequence typing

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Hui Zhang ◽  
Yuanyuan Wang ◽  
Lu Gao ◽  
Yan Wang ◽  
Rong Wei
Keyword(s):  
Multilocus Sequence Typing ◽  
Clinical Signs ◽  
Mycoplasma Hyopneumoniae ◽  
Rrna Gene ◽  
Genotype Diversity ◽  
Swine Diseases ◽  
Swine Herds ◽  
Pcr Targeting ◽  
Sequence Types ◽  
The 16S Rrna Gene

Abstract Background Between 2018 and 2020, 989 clinical specimens from pigs showing clinical signs of a variety of swine diseases in 27 provinces in China were sampled and submitted for further testing. Nested PCR targeting the 16S rRNA gene of Mycoplasma hyopneumoniae and subsequent sequencing were used to analyse these specimens. Mycoplasma hyopneumoniae-positive samples were assayed by multilocus sequence typing (MLST). The aim of the study was to reveal the distribution of M. hyopneumoniae and determine the genotypes of M. hyopneumoniae in pig herds in China based on MLST. Results Among these 989 samples, 199 samples were M. hyopneumoniae-positive. The M. hyopneumoniae positivity rate was 7.2% (35/494) in 2018, 18.4% (38/207) in 2019, and 43.8% (126/288) in 2020. In total, 47 samples were successfully assayed by MLST. Sixteen new M. hyopneumoniae sequence types from 9 provinces were recorded in the present study. Conclusions This is the first report on sample positivity rates and molecular typing results for M. hyopneumoniae in swine herds in China. MLST has revealed high genotype diversity among M. hyopneumoniae from different provinces of China.

scholarly journals Genetic diversity of Ehrlichia canisstrains from naturally infected dogs in Rio de Janeiro, Brazil

2014 ◽  
Vol 23 (3) ◽  
pp. 301-308 ◽  
Author(s):  
Renata Fernandes Ferreira ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Tatiana Xavier de Castro ◽  
Eliane de Oliveira Ferreira ◽  
Felipe Piedade Gonçalves Neves ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Rio De Janeiro ◽  
Complete Sequence ◽  
Rrna Gene ◽  
Hybrid Strain ◽  
The Us ◽  
Polymerase Chain ◽  
Hematological Manifestations ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

scholarly journals Molecular Detection of Borrelia Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity

Diagnostics ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 2088
Author(s):  
Malin Lager ◽  
Peter Wilhelmsson ◽  
Andreas Matussek ◽  
Per-Eric Lindgren ◽  
Anna J. Henningsson
Keyword(s):  
Cerebrospinal Fluid ◽  
16S Rrna ◽  
Clinical Diagnostics ◽  
Systematic Evaluation ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Analytical Sensitivity ◽  
Gene 16S Rrna ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

The main tools for clinical diagnostics of Lyme neuroborreliosis (LNB) are based on serology, i.e., detection of antibodies in cerebrospinal fluid (CSF). In some cases, PCR may be used as a supplement, e.g., on CSF from patients with early LNB. Standardisation of the molecular methods and systematic evaluation of the pre-analytical handling is lacking. To increase the analytical sensitivity for detection of Borrelia bacteria in CSF by PCR targeting the 16S rRNA gene, parameters were systematically evaluated on CSF samples spiked with a known amount of cultured Borrelia bacteria. The results showed that the parameters such as centrifugation time and speed, the use of complementary DNA as a template (in combination with primers and a probe aiming at target gene 16S rRNA), and the absence of inhibitors (e.g., erythrocytes) had the highest impact on the analytical sensitivity. Based on these results, a protocol for optimised handling of CSF samples before molecular analysis was proposed. However, no clinical evaluation of the proposed protocol has been done so far, and further investigations of the diagnostic sensitivity need to be performed on well-characterised clinical samples from patients with LNB.

scholarly journals Accuracy of Conventional PCR Targeting the 16S rRNA Gene with the Ot-16sRF1 and Ot-16sRR1 Primers for Diagnosis of Scrub Typhus: a Case-Control Study: TABLE 1

2015 ◽  
Vol 54 (1) ◽  
pp. 178-179 ◽  
Author(s):  
Choon-Mee Kim ◽  
Min Keun Cho ◽  
Dong-Min Kim ◽  
Na-Ra Yun ◽  
Seok Won Kim ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Scrub Typhus ◽  
Rrna Gene ◽  
Conventional Pcr ◽  
Content Type ◽  
Increased Sensitivity ◽  
Good Diagnostic Accuracy ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

We retrospectively evaluated the accuracy of conventional PCR targeting the 16S rRNA gene (16S C-PCR) using the Ot-16sRF1/Ot-16sRR1 primers for diagnosing scrub typhus. The diagnosis ofOrientia tsutsugamushiinfection by 16S C-PCR presented an increased sensitivity of 87.0% and specificity of 100% compared with those obtained with other targets and is thus a simple and clinically useful method with good diagnostic accuracy.

scholarly journals Development of a Multilocus Sequence Typing Tool for High-Resolution Genotyping of Enterocytozoon bieneusi

2011 ◽  
Vol 77 (14) ◽  
pp. 4822-4828 ◽  
Author(s):  
Yaoyu Feng ◽  
Na Li ◽  
Theresa Dearen ◽  
Maria L. Lobo ◽  
Olga Matos ◽  
...  
Keyword(s):  
High Resolution ◽  
Multilocus Sequence Typing ◽  
Enterocytozoon Bieneusi ◽  
Whole Genome Sequence ◽  
Rrna Gene ◽  
Content Type ◽  
Health Significance ◽  
The One ◽  
Pcr Targeting ◽  
Typing Tool

ABSTRACTThus far, genotyping ofEnterocytozoon bieneusihas been based solely on DNA sequence analysis of the internal transcribed spacer (ITS) of the rRNA gene. Both host-adapted and zoonotic (human-pathogenic) genotypes ofE. bieneusihave been identified. In this study, we searched for microsatellite and minisatellite sequences in the whole-genome sequence database ofE. bieneusiisolate H348. Seven potential targets (MS1 to MS7) were identified. Testing of the seven targets by PCR using two human-pathogenicE. bieneusigenotypes (A and Peru10) led to the selection of four targets (MS1, MS3, MS4, and MS7). Further analysis of the four loci with an additional 24 specimens of both host-adapted and zoonoticE. bieneusigenotypes indicated that most host-adapted genotypes were not amplified by PCR targeting these loci. In contrast, 10 or 11 of the 13 specimens of the zoonotic genotypes were amplified by PCR at each locus. Altogether, 12, 8, 7, and 11 genotypes of were identified at MS1, MS3, MS4, and MS7, respectively. Phylogenetic analysis of the nucleotide sequences obtained produced a genetic relationship that was similar to the one at the ITS locus, with the formation of a large group of zoonotic genotypes that included mostE. bieneusigenotypes in humans. Thus, a multilocus sequence typing tool was developed for high-resolution genotyping ofE. bieneusi.Data obtained in the study should also have implications for understanding the taxonomy ofEnterocytozoonspp., the public health significance ofE. bieneusiin animals, and the sources of humanE. bieneusiinfections.

scholarly journals Detection of Anaplasma sp. phylogenetically related to A. phagocytophilum in a free-living bird in Brazil

2017 ◽  
Vol 26 (4) ◽  
pp. 505-510 ◽  
Author(s):  
Anna Claudia Baumel Mongruel ◽  
Jyan Lucas Benevenute ◽  
Priscila Ikeda ◽  
Marcos Rogério André ◽  
Rosangela Zacarias Machado ◽  
...  
Keyword(s):  
Wild Bird ◽  
Molecular Detection ◽  
Bird Species ◽  
Rrna Gene ◽  
Wild Animals ◽  
Domestic Cats ◽  
Free Living ◽  
Likelihood Analysis ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

Abstract Wild animals play an important role in carrying vectors that may potentially transmit pathogens. Several reports highlighted the participation of wild animals on the Anaplasma phagocytophilum cycle, including as hosts of the agent. The aim of this study was to report the molecular detection of an agent phylogenetically related to A. phagocytophilum isolated from a wild bird in the Midwest of the state of Paraná, Brazil. Fifteen blood samples were collected from eleven different bird species in the Guarapuava region. One sample collected from a Penelope obscura bird was positive in nested PCR targeting the 16S rRNA gene of Anaplasma spp. The phylogenetic tree based on the Maximum Likelihood analysis showed that the sequence obtained was placed in the same clade with A. phagocytophilum isolated from domestic cats in Brazil. The present study reports the first molecular detection of a phylogenetically related A. phagocytophilum bacterium in a bird from Paraná State.

scholarly journals Overestimation of the Abundance of Sulfate-Reducing Bacteria in Human Feces by Quantitative PCR Targeting the Desulfovibrio 16S rRNA Gene

2011 ◽  
Vol 77 (10) ◽  
pp. 3544-3546 ◽  
Author(s):  
C. T. Christophersen ◽  
M. Morrison ◽  
M. A. Conlon
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Quantitative Pcr ◽  
Sulfate Reducing Bacteria ◽  
Rrna Gene ◽  
Sulfate Reducing ◽  
Gene Assay ◽  
Reducing Bacteria ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

ABSTRACTThe dominant genus of sulfate-reducing bacteria (SRB) in humans isDesulfovibrio, and quantitative PCR (QPCR) targeting the 16S rRNA gene is often used in assays. We show that the 16S rRNA gene assay overestimated SRB abundance in feces from 24 adults compared to QPCR assays using primers targeting two genes involved in SRB energy metabolism.

scholarly journals Chlamydiales and hemotropic mycoplasma in captive and free-living bats

2020 ◽  
Author(s):  
Janine Fritschi ◽  
Hanna Marti ◽  
Helena M.B. Seth-Smith ◽  
Sébastien Aeby ◽  
Gilbert Greub ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Rrna Gene ◽  
Free Living ◽  
Hemotropic Mycoplasma ◽  
Total Prevalence ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

Abstract Background: Bats are hosts for a variety of microorganisms, however, little is known about the presence of Chlamydiales and hemotropic mycoplasma. This study investigated 475 free-living and captive bats from Switzerland, Germany and Costa Rica for the occurrence of Chlamydiales and hemotropic mycoplasma.Results: Screening for Chlamydiales was performed using a Chlamydiaceae-specific real-time PCR targeting the 23S rRNA gene and a pan-Chlamydiales PCR targeting the 16S rRNA gene resulting in a total prevalence of 31.4%. For sequencing, a PCR with the specifically designed inner primers panFseq and panRseq was performed, and criteria published by Pillonel et al. were used to classify the 19 obtained sequences, resulting in the formation of two groups. Groups one and two shared sequence identities to Chlamydiaceae and to Chlamydia-like organisms, including Rhabdochlamydiaceae and unclassified Chlamydiales from environmental samples, respectively.Analysis for the presence of hemotropic mycoplasma was performed using a universal SYBR Green hemoplasma screening real-time PCR targeting the 16S rRNA gene, real-time PCRs specific for M. haemofelis-like and 'Candidatus M. haemominutum'-like organisms and two conventional PCRs targeting an 871-bp and 1030-bp region of the 16S rRNA gene resulting in a total prevalence of 0.7%. Sequencing and phylogenetic analysis of the 871-bp and 1030-bp region of the 16S rRNA gene were used to classify positive specimens and infer their phylogenetic relationships. Three sequences with identities to other unidentified mycoplasma found in vampire bats and Chilean bats were obtained.Conclusions: Bats can harbor Chlamydiales and hemotropic mycoplasma and the newly described sequences in this study indicate that the diversity of these bacteria in bats is much larger than thought before. Both, Chlamydiales and hemotropic mycoplasmas are not restricted to certain bat species or countries and free-living as well as captive bats can be colonized. In conclusion, bats represent another potential host or vector for novel, previously unidentified, Chlamydiales and hemotropic mycoplasma.

scholarly journals Competitive PCR for Quantitation of a Cytophaga-Flexibacter-Bacteroides Phylum Bacterium Associated with the Tuber borchii Vittad. Mycelium

2002 ◽  
Vol 68 (12) ◽  
pp. 6421-6424 ◽  
Author(s):  
Elena Barbieri ◽  
Giulia Riccioni ◽  
Anna Pisano ◽  
Davide Sisti ◽  
Sabrina Zeppa ◽  
...  
Keyword(s):  
16S Rrna ◽  
Mycorrhizal Fungus ◽  
Ectomycorrhizal Fungus ◽  
Rrna Gene ◽  
Competitive Pcr ◽  
Tuber Borchii ◽  
Uncultured Bacterium ◽  
Pcr Targeting ◽  
The 16S Rrna Gene ◽  
Number Of Bacteria

ABSTRACT An uncultured bacterium associated with the ectomycorrhizal fungus Tuber borchii Vittad. was identified as a novel member of the Cytophaga-Flexibacter-Bacteroides group. Utilizing a quantitative PCR targeting the 16S rRNA gene, we relatively quantified this bacterium in the host. The estimated number of bacteria was found to be approximately 106 cells per 30-day-old T. borchii mycelium culture. This represents the first molecular attempt to enumerate an uncultured bacterium associated with a mycorrhizal fungus.

First molecular survey and identification ofAnaplasmaspp. in white yaks (Bos grunniens) in China

Parasitology ◽  
2016 ◽  
Vol 143 (6) ◽  
pp. 686-691 ◽  
Author(s):  
JIFEI YANG ◽  
ZHIJIE LIU ◽  
QINGLI NIU ◽  
JUNLONG LIU ◽  
GUIQUAN GUAN ◽  
...  
Keyword(s):  
Northwest China ◽  
Intracellular Bacteria ◽  
Rrna Gene ◽  
Obligate Intracellular ◽  
Obligate Intracellular Bacteria ◽  
Control And Management ◽  
Sequence Types ◽  
Anaplasma Bovis ◽  
The 16S Rrna Gene

SUMMARYAnaplasmosis is caused by a group of obligate intracellular bacteria in the genusAnaplasma, which are transmitted by ticks and infect humans, domestic animals and wildlife. This study was conducted to determine the prevalence and molecular characterization ofAnaplasmaspp. in semi-wild white yaks sampled in Tianzhu Tibetan Autonomous County, northwest China. Out of 332 samples tested, 35 (10·9%) were positive forAnaplasmaspp. The positive rates were 6·2% (20/322) and 5·3% (17/322) forAnaplasma bovisandAnaplasma phagocytophilumin white yaks, respectively. None of the sample was positive forAnaplasma marginale. Two (0·6%) samples were simultaneously positive toA. bovisandA. phagocytophilum. Sequence analysis of the 16S rRNA gene revealed two genotypes (ApG1 and ApG2) ofA. phagocytophilumand two sequence types (ST1 and ST2) ofA. bovisin white yaks. This study is the first to document the presence ofAnaplasmain white yaks. Our findings extend the host range forAnaplasmaspecies and provide more valuable information for the control and management of anaplasmosis in white yaks.

Sign in / Sign up

Export Citation Format

Share Document