Molecular Evidence for Local Acquisition of Human Alveolar Echinococcosis in Saskatchewan, Canada

2020 ◽  
Author(s):  
Janna M Schurer ◽  
Polly Tsybina ◽  
Karen M Gesy ◽  
Temitope U Kolapo ◽  
Stuart Skinner ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Haplotype Analysis ◽  
Alveolar Echinococcosis ◽  
Mitochondrial Gene ◽  
Molecular Evidence ◽  
Travel History ◽  
Life Threatening ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Autochthonous Transmission

Abstract Alveolar echinococcosis (AE) is a life-threatening parasitic disease caused by the zoonotic cestode Echinococcus multilocularis. Our goals were to confirm infection, identify species, and analyze biogeographical origin of metacestode tissues from a suspected human AE case in Saskatchewan, Canada. We conducted polymerase chain reaction (PCR) targeting the nad1 mitochondrial gene for E. multilocularis and the rrnS ribosomal RNA gene for E. granulosus and conducted haplotype analysis at the nad2 locus. Our analysis confirmed AE and indicated that sequences matched infected Saskatchewan coyotes and European E3/E4 haplotypes. The patient had no travel history outside North America. This suggests autochthonous transmission of a European-type strain.

scholarly journals Molecular Identification of Trypanosoma evansi Isolated from Arabian Camels (Camelus dromedarius) in Riyadh and Al-Qassim, Saudi Arabia

Animals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 1149
Author(s):  
Dina M. Metwally ◽  
Isra M. Al-Turaiki ◽  
Najwa Altwaijry ◽  
Samia Q. Alghamdi ◽  
Abdullah D. Alanazi
Keyword(s):  
Saudi Arabia ◽  
Infection Rate ◽  
Ribosomal Rna ◽  
Phylogenetic Analyses ◽  
Trypanosoma Evansi ◽  
Camelus Dromedarius ◽  
18S Ribosomal Rna ◽  
18S Ribosomal Rna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting

We analyzed the blood from 400 one-humped camels, Camelus dromedarius (C. dromedarius), in Riyadh and Al-Qassim, Saudi Arabia to determine if they were infected with the parasite Trypanosoma spp. Polymerase chain reaction (PCR) targeting the internal transcribed spacer 1 (ITS1) gene was used to detect the prevalence of Trypanosoma spp. in the camels. Trypanosoma evansi (T. evansi) was detected in 79 of 200 camels in Riyadh, an infection rate of 39.5%, and in 92 of 200 camels in Al-Qassim, an infection rate of 46%. Sequence and phylogenetic analyses revealed that the isolated T. evansi was closely related to the T. evansi that was detected in C. dromedarius in Egypt and the T. evansi strain B15.1 18S ribosomal RNA gene identified from buffalo in Thailand. A BLAST search revealed that the sequences are also similar to those of T. evansi from beef cattle in Thailand and to T. brucei B8/18 18S ribosomal RNA from pigs in Nigeria.

scholarly journals Detecting Plasmodium falciparum in community surveys: a comparison of Paracheck Pf® Test and ICT Malaria Pf® Cassette Test to polymerase chain reaction in Mutasa District, Zimbabwe

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Nobert Mudare ◽  
Zvifadzo Matsena-Zingoni ◽  
Aramu Makuwaza ◽  
Edmore Mamini ◽  
Shungu S. Munyati ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Falciparum ◽  
Mitochondrial Gene ◽  
Dried Blood Spots ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Cross Sectional ◽  
Predictive Values ◽  
Polymerase Chain ◽  
Pcr Targeting

Abstract Background Microscopy and rapid diagnostic tests (RDTs) are the main techniques used to diagnose malaria. While microscopy is considered the gold standard, RDTs have established popularity as they allow for rapid diagnosis with minimal technical skills. This study aimed to compare the diagnostic performance of two Plasmodium falciparum histidine-rich protein 2 (PfHRP2)-based RDTs (Paracheck Pf® Test (Paracheck) and Malaria Pf™ ICT (ICT)) to polymerase chain reaction (PCR) in a community survey. Methods A cross-sectional study was conducted between October 2012 and December 2014 in Mutasa District, Manicaland Province, eastern Zimbabwe. Households were randomly selected using satellite imagery, and 224 households were visited. Residents present in the household on the date of the visit were recruited for the study. Participants of all age groups from the selected households were screened with Paracheck and ICT RDTs in parallel. Dried blood spots (DBS) and thin and thick smears were collected. Parasite DNA extracted from the DBS was subjected to nested PCR targeting the Plasmodium cytochrome b mitochondrial gene. Data analysis was performed using the Cohen’s Kappa test to determine the interrater agreement and the sensitivity and specificity of the diagnostic test were reported. Results Results from a total of 702 participants were analysed. Most were females, 397 (57%), and the median age of participants was 21 years with an interquartile range of 9–39 years. Of those who were screened, 8 (1.1%), 35 (5.0%), and 21 (2.9%) were malaria parasite positive by microscopy, RDT and PCR, respectively. Paracheck and ICT RDTs had a 100% agreement. Comparing RDT and PCR results, 34 participants (4.8%) had discordant results. Most of the discordant cases were RDT positive but PCR negative (n = 24). Half of those RDT positive, but PCR negative individuals reported anti-malarials to use in the past month, which is significantly higher than reported anti-malarial drug use in the population (p < 0.001). The participant was febrile on the day of the visit, but relying on PfHRP2-based RDT would miss this case. Among the diagnostic methods evaluated, with reference to PCR, the sensitivity was higher with the RDT (52.4%) while specificity was higher with the microscopy (99.9%). The positive predictive value (PPV) was higher with the microscopy (87.5%), while the negative predictive values were similar for both microscopy and RDTs (98%). Overall, a strong correlated agreement with PCR was observed for the microscopy (97.9%) and the RDTs (95.2%). Conclusions Paracheck and ICT RDTs showed 100% agreement and can be used interchangeably. As malaria transmission declines and Zimbabwe aims to reach malaria elimination, management of infected individuals with low parasitaemia as well as non-P. falciparum infection can be critical.

scholarly journals Molecular confirmation of hemothropic mycoplasmas (hemoplasmas) in domestic cats (Felis catus) in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Viorel Herman ◽  
...  
Keyword(s):  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Candidatus Mycoplasma Turicensis ◽  
Sampling Season

Abstract Background: The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods : Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results: Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0 . 05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions : The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

Molecular evidence of Francisella noatunensis sub sp. orientalis in Oreochromis niloticus in Yunnan Province, south-western China

2020 ◽  
Vol 67 (2) ◽  
Author(s):  
Guoyi Niu ◽  
Rutch Khattiya ◽  
Sukolrat Boonyayatra ◽  
Dilok Wongsathein ◽  
William Hueston
Keyword(s):  
Ribosomal Rna ◽  
Oreochromis Niloticus ◽  
Yunnan Province ◽  
Management Practices ◽  
Bacterial Pathogen ◽  
Quality Parameters ◽  
Western China ◽  
Molecular Evidence ◽  
Granulomatous Lesions ◽  
Polymerase Chain

Francisella sp. is an emerging bacterial pathogen affecting tilapia Oreochromis niloticus This study surveyed presence of Francisella sp. in tilapia raised in Yunnan Province, south-western China along with water quality parameters and management practices. A total of 224 tilapia specimens were sampled from 28 farms between June and September 2017. Necropsy of the sampled fish revealed no granulomatous lesions and the standard bacterial isolation protocols proved negative for Francisella sp. However, the presence of Francisella sp. was confirmed by polymerase chain reaction in 18 samples (8%) from 8 of the 28 farms (28.6%) sampled, though no increasing mortality was reported from these farms. The 16S ribosomal RNA gene sequences revealed 99.7% identity with F. noatunensis sub sp. orientalis. The presence of Francisella sp. infection showed significant association with water temperature.

scholarly journals Molecular confirmation of hemothropic mycoplasmas (hemoplasmas) in domestic cats in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Tijana Suici ◽  
...  
Keyword(s):  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Candidatus Mycoplasma Turicensis ◽  
Sampling Season

Abstract Background The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Abundant Mitochondrial Genome Diversity, Population Differentiation and Convergent Evolution in Pines

Genetics ◽  
1998 ◽  
Vol 150 (4) ◽  
pp. 1605-1614
Author(s):  
Junyuan Wu ◽  
Konstantin V Krutovskii ◽  
Steven H Strauss
Keyword(s):  
Mitochondrial Dna ◽  
Population Differentiation ◽  
Mitochondrial Gene ◽  
Dna Polymorphisms ◽  
Neighbor Joining ◽  
Closely Related Species ◽  
Breeding Programs ◽  
Length Polymorphisms ◽  
Polymerase Chain ◽  
Mitochondrial Genome Diversity

Abstract We examined mitochondrial DNA polymorphisms via the analysis of restriction fragment length polymorphisms in three closely related species of pines from western North America: knobcone (Pinus attenuata Lemm.), Monterey (P. radiata D. Don), and bishop (P. muricata D. Don). A total of 343 trees derived from 13 populations were analyzed using 13 homologous mitochondrial gene probes amplified from three species by polymerase chain reaction. Twenty-eight distinct mitochondrial DNA haplotypes were detected and no common haplotypes were found among the species. All three species showed limited variability within populations, but strong differentiation among populations. Based on haplotype frequencies, genetic diversity within populations (HS) averaged 0.22, and population differentiation (GST and θ) exceeded 0.78. Analysis of molecular variance also revealed that &gt;90% of the variation resided among populations. For the purposes of genetic conservation and breeding programs, species and populations could be readily distinguished by unique haplotypes, often using the combination of only a few probes. Neighbor-joining phenograms, however, strongly disagreed with those based on allozymes, chloroplast DNA, and morphological traits. Thus, despite its diagnostic haplotypes, the genome appears to evolve via the rearrangement of multiple, convergent subgenomic domains.

Frequency of the delta Phe508 mutation and correlation with XV.2c/KM-19 haplotypes in an American population of cystic fibrosis patients: results of a collaborative study

1990 ◽  
Vol 36 (10) ◽  
pp. 1741-1746 ◽  
Author(s):  
W E Highsmith ◽  
G L Chong ◽  
H T Orr ◽  
T R Perry ◽  
D Schald ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Haplotype Analysis ◽  
Screening Program ◽  
Collaborative Study ◽  
Population Based ◽  
Carrier Status ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Phenylalanine Residue ◽  
Test Result

Abstract The cystic fibrosis (CF) gene has been recently cloned, and a deletion of 3 basepairs (bp) of DNA was found on most of the CF chromosomes. This deletion leads to the synthesis of a protein that lacks a phenylalanine residue at position 508. Using two polymerase chain reaction protocols to study the frequency of this mutation in a series of 192 CF patients, we found the mutation on 72% of affected chromosomes. We then used this value to calculate the predictive value of a negative test result in a population-based screening program for CF carrier status. Haplotype analysis with the polymorphic markers XV.2c and KM-19 on 239 CF chromosomes revealed that 90.7% of CF chromosomes with the deletion had a single haplotype. This haplotype was also associated with 60.4% of CF chromosomes with unknown mutations. These values can be used to calculate the probability of whether an individual from the general population is a carrier of any CF mutation.

Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides

2021 ◽  
pp. 030098582199156
Author(s):  
Alexandra N. Myers ◽  
Unity Jeffery ◽  
Zachary G. Seyler ◽  
Sara D. Lawhon ◽  
Aline Rodrigues Hoffmann
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Accuracy ◽  
Molecular Techniques ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Identification Of Fungi ◽  
Fungal Dna ◽  
Polymerase Chain ◽  
Panfungal Pcr ◽  
Pcr Targeting

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

Lemierre Syndrome: Not So Forgotten!

2014 ◽  
Vol 23 (2) ◽  
pp. 176-179 ◽  
Author(s):  
Nishant Gupta ◽  
Stephen M. Kralovic ◽  
Dennis McGraw
Keyword(s):  
Internal Jugular Vein ◽  
Jugular Vein ◽  
Lymphatic Vessels ◽  
Fusobacterium Necrophorum ◽  
Septic Thrombophlebitis ◽  
Lemierre Syndrome ◽  
Travel History ◽  
Life Threatening Illness ◽  
Life Threatening ◽  
Bedside Ultrasonography

Lemierre syndrome is a rare and life-threatening illness. Often referred to as “the forgotten disease,” its incidence is reported to be as low as 1 in a million. The microorganism responsible for Lemierre syndrome is typically Fusobacterium necrophorum. The bacterium starts in the pharynx and peritonsillar tissue, then disseminates through lymphatic vessels. Severe sepsis rapidly develops, as does the hallmark of this syndrome: septic thrombophlebitis of the internal jugular vein. This report describes a case of Lemierre syndrome in a previously healthy 26-year-old man with life-threatening internal jugular vein thrombophlebitis following 2 weeks of an indolent course of pharyngitis. The patient’s initial presentation and extensive travel history as an Army veteran were particularly challenging aspects in establishing his diagnosis. The diagnosis of Lemierre syndrome is frequently delayed. Routine use of bedside ultrasonography may aid in rapid diagnosis of the disease.

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