scholarly journals Pathogen Detection and Genotyping With Trace Sample by A Phi29 Based Unbiased Exponential Amplification

2021 ◽  
Author(s):  
Xi Zhang ◽  
Jingjing Chen ◽  
Heling Xu ◽  
Qi Zhang ◽  
Huan Zhang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Northeast China ◽  
Pathogen Detection ◽  
Spotted Fever ◽  
Liaoning Province ◽  
Growth Stages ◽  
Spotted Fever Group ◽  
New Genotype ◽  
Pathogen Screening ◽  
Multiple Pathogen

Abstract Ticks are vectors for many infectious diseases such as spotted fever group (SFG) rickettsioses and borrelioses. Ticks are valuable material for pathogen ecology study. Ticks have several growth stages with significant varying size, and therefore, in most cases, the collected ticks cannot provide sufficient DNA for subsequent studies, particularly for multiple pathogen screening and genotyping. Unbiased pretreatment of the tick samples for subsequent analysis is an urgent need for subsequent ecological survey and other studies. Phi29 DNA polymerase, an enzyme with strand displacement activity, could exponentially amplify DNA randomly and non-biasedly, generating large quantities of DNA. In the present study, we developed a Phi29 based unbiased exponential amplification (PEA) assay for unbiased treatment of sample nucleic acid to provide sufficient DNA for genetic analysis. By using tick borne pathogen detection and genotype as a model, we tested and evaluated the feasibility of the assay. Nucleic acid were extracted from single ticks and subjected to PEA. The results showed that tick DNA could be amplified up to 10 5 folds. The amplified products were successfully used for pathogen screening and genotyping. With the amplified DNA from single tick, Rickettsia was successfully detected and genotyped. A new genotype of Rickettsia was identified from ticks collected from Dandong city, Liaoning province, Northeast China. This PEA assay is universal and can also be extended to other applications where samples are greatly limited.

scholarly journals Detection of tick-borne pathogens in questing Ixodes ricinus in the French Pyrenees and first identification of Rickettsia monacensis in France

Parasite ◽  
2019 ◽  
Vol 26 ◽  
pp. 20 ◽  
Author(s):  
Toufic Akl ◽  
Gilles Bourgoin ◽  
Marie-Line Souq ◽  
Joël Appolinaire ◽  
Marie-Thérèse Poirel ◽  
...  
Keyword(s):  
Ixodes Ricinus ◽  
Pathogen Detection ◽  
Spotted Fever ◽  
Spotted Fever Group ◽  
Rupicapra Pyrenaica ◽  
Rickettsia Monacensis ◽  
Animal Pathogens ◽  
Questing Ticks ◽  
Pyrenean Chamois ◽  
First Time

Ticks are important vectors of several human and animal pathogens. In this study, we estimated the prevalence of important tick-borne infections in questing ticks from an area in Southwestern France (Hautes-Pyrénées) inhabited by Pyrenean chamois (Rupicapra pyrenaica pyrenaica) experiencing high tick burden. We examined adult and nymph ticks collected by the flag dragging method from 8 to 15 sites in the Pic de Bazès during the years 2009, 2011, 2013 and 2015. PCR assays were conducted on selected ticks for the detection of Borrelia burgdorferi s.l., Babesia spp., Rickettsia spp., spotted fever group (SFG) Rickettsia and Anaplasma phagocytophilum. Randomly selected positive samples were submitted for sequence analysis. A total of 1971 questing ticks were collected including 95 males, 101 females and 1775 nymphs. All collected ticks were identified as Ixodes ricinus. Among them, 696 ticks were selected for pathogen detection and overall prevalence was 8.4% for B. burgdorferi s.l.; 0.4% for Babesia spp.; 6.1% for A. phagocytophilum; 17.6% for Rickettsia spp.; and 8.1% for SFG Rickettsia. Among the sequenced pathogens, we detected in this population of ticks the presence of Babesia sp. EU1 and Rickettsia helvetica, as well as Rickettsia monacensis for the first time in France. The detection of these pathogens in the Pic de Bazès highlights the potential infection risks for visitors to this area and the Pyrenean chamois population.

Rickettsiae of the spotted-fever group in ixodid ticks from Hungary: identification of a new genotype ('Candidatus Rickettsia kotlanii')

2006 ◽  
Vol 100 (3) ◽  
pp. 229-236 ◽  
Author(s):  
Z. Sréter-Lancz ◽  
Z. Széll ◽  
G. Kovács ◽  
L. Egyed ◽  
K. Márialigeti ◽  
...  
Keyword(s):  
Spotted Fever ◽  
Ixodid Ticks ◽  
Spotted Fever Group ◽  
Genotype I ◽  
New Genotype

scholarly journals Two Cases of Spotted Fever Group Rickettsiosis Contracted in Southern Parts of Africa

1998 ◽  
Vol 72 (12) ◽  
pp. 1311-1316 ◽  
Author(s):  
Mikio KIMURA ◽  
Tatsuya FUJII ◽  
Aikichi IWAMOTO
Keyword(s):  
Spotted Fever ◽  
Spotted Fever Group

scholarly journals Surveillance of Ticks and Tick-Borne Pathogens in Suburban Natural Habitats of Central Maryland

2021 ◽  
Author(s):  
Matthew T Milholland ◽  
Lars Eisen ◽  
Robyn M Nadolny ◽  
Andrias Hojgaard ◽  
Erika T Machtinger ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Spotted Fever ◽  
Babesia Microti ◽  
Borrelia Burgdorferi Sensu Lato ◽  
Spotted Fever Group ◽  
Borrelia Miyamotoi ◽  
Natural Habitats ◽  
Questing Ticks ◽  
Tick Vectors

Abstract Lyme and other tick-borne diseases are increasing in the eastern United States and there is a lack of research on integrated strategies to control tick vectors. Here we present results of a study on tick-borne pathogens detected from tick vectors and rodent reservoirs from an ongoing 5-yr tick suppression study in the Lyme disease-endemic state of Maryland, where human-biting tick species, including Ixodes scapularis Say (Acari: Ixodidae) (the primary vector of Lyme disease spirochetes), are abundant. During the 2017 tick season, we collected 207 questing ticks and 602 ticks recovered from 327 mice (Peromyscus spp. (Rodentia: Cricetidae)), together with blood and ear tissue from the mice, at seven suburban parks in Howard County. Ticks were selectively tested for the presence of the causative agents of Lyme disease (Borrelia burgdorferi sensu lato [s.l.]), anaplasmosis (Anaplasma phagocytophilum), babesiosis (Babesia microti), ehrlichiosis (Ehrlichia ewingii, Ehrlichia chaffeensis, and ‘Panola Mountain’ Ehrlichia) and spotted fever group rickettsiosis (Rickettsia spp.). Peromyscus ear tissue and blood samples were tested for Bo. burgdorferi sensu stricto (s.s), A. phagocytophilum, Ba. microti, and Borrelia miyamotoi. We found 13.6% (15/110) of questing I. scapularis nymphs to be Bo. burgdorferi s.l. positive and 1.8% (2/110) were A. phagocytophilum positive among all sites. Borrelia burgdorferi s.s. was found in 71.1% (54/76) of I. scapularis nymphs removed from mice and 58.8% (194/330) of captured mice. Results from study on tick abundance and pathogen infection status in questing ticks, rodent reservoirs, and ticks feeding on Peromyscus spp. will aid efficacy evaluation of the integrated tick management measures being implemented.

Multifunctional Printable Micropattern Array for Digital Nucleic Acid Assay for Microbial Pathogen Detection

2021 ◽  
Vol 13 (2) ◽  
pp. 3098-3108
Author(s):  
Yunho Choi ◽  
Younseong Song ◽  
Yong Tae Kim ◽  
Seok Jae Lee ◽  
Kyoung G. Lee ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Pathogen Detection ◽  
Microbial Pathogen ◽  
Nucleic Acid Assay

scholarly journals Isolation of spotted fever group rickettsiae from Ixodes persulcatus ticks in the central part of Hokkaido, Japan

2000 ◽  
Vol 51 (1) ◽  
pp. 55-58 ◽  
Author(s):  
Hiromi FUJITA ◽  
Nobuhiro TAKADA ◽  
Emiko ISOGAI ◽  
Yuriko WATANABE ◽  
Takuya ITO
Keyword(s):  
Spotted Fever ◽  
Ixodes Persulcatus ◽  
Spotted Fever Group ◽  
Spotted Fever Group Rickettsiae

scholarly journals Quantitative Analyses of Variations in the Injury of Endothelial Cells Elicited by 11 Isolates of Rickettsia rickettsii

2001 ◽  
Vol 8 (4) ◽  
pp. 788-796 ◽  
Author(s):  
Marina E. Eremeeva ◽  
Gregory A. Dasch ◽  
David J. Silverman
Keyword(s):  
North Carolina ◽  
Endothelial Cells ◽  
Reduced Glutathione ◽  
Spotted Fever ◽  
Ixodid Ticks ◽  
Spotted Fever Group ◽  
Cellular Injury ◽  
Rickettsia Rickettsii ◽  
Cytopathic Effects ◽  
Group I

ABSTRACT Eleven isolates of spotted fever group rickettsiae from the blood of patients or ixodid ticks from North and South America were characterized. All isolates were identified as Rickettsia rickettsii using restriction fragment length polymorphism analysis of a 532-bp rOmpA gene fragment obtained by PCR. The ability of the R. rickettsii isolates to elicit cytopathic effects and parameters of oxidative injury were examined in cultured human EA.hy 926 endothelial cells. Cytopathic effects were determined by direct observation of infected cultures, by measuring the release of cytoplasmic lactate dehydrogenase (LDH), and by determination of intracellular pools of peroxide and reduced glutathione. Four biotypes of R. rickettsii were defined. Group I included two highly cytopathic isolates from Montana, Bitterroot and Sheila Smith, and three isolates from Maryland, North Carolina, and Brazil. These isolates rapidly damaged cells, released large amounts of cytoplasmic LDH, caused accumulation of intracellular peroxide, and depleted intracellular pools of reduced glutathione. Group II contained three isolates, two from Montana, Hlp#2 and Lost Horse Canyon, and an isolate from Colombia, which were similar to group I but caused either lower responses in LDH release or smaller changes in intracellular peroxide levels. The group III isolates, Sawtooth from Montana and 84JG from North Carolina, caused lower cellular injury by all measures. Group IV isolate Price T from Montana was the least cytopathic and caused minimal alterations of all parameters measured. Understanding the molecular basis for the varied cellular injury caused by different isolates of R. rickettsii may contribute to improved treatment of Rocky Mountain spotted fever and to the rapid identification of those isolates which are more likely to cause fulminant disease.

scholarly journals Seroepidemiologic Study of Human Infections with Spotted Fever Group Rickettsiae in North Carolina

2014 ◽  
Vol 52 (11) ◽  
pp. 3960-3966 ◽  
Author(s):  
M. F. Vaughn ◽  
J. Delisle ◽  
J. Johnson ◽  
G. Daves ◽  
C. Williams ◽  
...  
Keyword(s):  
North Carolina ◽  
Spotted Fever ◽  
Spotted Fever Group ◽  
Spotted Fever Group Rickettsiae ◽  
Human Infections ◽  
Seroepidemiologic Study

scholarly journals Significant Growth by Rickettsia Species within Human Macrophage-Like Cells Is a Phenotype Correlated with the Ability to Cause Disease in Mammals

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 228
Author(s):  
M. Nathan Kristof ◽  
Paige E. Allen ◽  
Lane D. Yutzy ◽  
Brandon Thibodaux ◽  
Christopher D. Paddock ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Spotted Fever ◽  
Growth Characteristic ◽  
Human Macrophage ◽  
Rickettsia Conorii ◽  
Spotted Fever Group ◽  
Rocky Mountain Spotted Fever ◽  
Phagocytic Cells ◽  
Pathogenic Potential ◽  
Rickettsia Species

Rickettsia are significant sources of tick-borne diseases in humans worldwide. In North America, two species in the spotted fever group of Rickettsia have been conclusively associated with disease of humans: Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, and Rickettsia parkeri, the cause of R. parkeri rickettsiosis. Previous work in our lab demonstrated non-endothelial parasitism by another pathogenic SFG Rickettsia species, Rickettsia conorii, within THP-1-derived macrophages, and we have hypothesized that this growth characteristic may be an underappreciated aspect of rickettsial pathogenesis in mammalian hosts. In this work, we demonstrated that multiple other recognized human pathogenic species of Rickettsia, including R. rickettsii, R. parkeri, Rickettsia africae, and Rickettsiaakari can grow within target endothelial cells as well as within PMA-differentiated THP-1 cells. In contrast, Rickettsia bellii, a Rickettsia species not associated with disease of humans, and R. rickettsii strain Iowa, an avirulent derivative of pathogenic R. rickettsii, could invade both cell types but proliferate only within endothelial cells. Further analysis revealed that similar to previous studies on R. conorii, other recognized pathogenic Rickettsia species could grow within the cytosol of THP-1-derived macrophages and avoided localization with two different markers of lysosomal compartments; LAMP-2 and cathepsin D. R. bellii, on the other hand, demonstrated significant co-localization with lysosomal compartments. Collectively, these findings suggest that the ability of pathogenic rickettsial species to establish a niche within macrophage-like cells could be an important factor in their ability to cause disease in mammals. These findings also suggest that analysis of growth within mammalian phagocytic cells may be useful to predict the pathogenic potential of newly isolated and identified Rickettsia species.

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