Comparison of Japanese Isolates of <i>Coxiella burnetii</i> by PCR‐RFLP and Sequence Analysis

Andoh, Masako; Nagaoka, Hiromi; Yamaguchi, Tsuyoshi; Fukushi, Hideto; Hirai, Katsuya

doi:10.1111/j.1348-0421.2004.tb03627.x

Cited by 12 publications

(8 citation statements)

References 23 publications

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“…Thus, certain plasmid patterns were claimed to be associated with the disease outcome [3,4], which was controversial [5]; also, some isocitrate dehydrogenase types were associated with chronic disease and a role for this gene in the adaptation of the organism to the intracellular environment was proposed [6], although this association was also challenged by other authors [7]. …”

Section: Introductionmentioning

confidence: 99%

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

et al. 2012

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BackgroundCoxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes.ResultsTo assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct characterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples.ConclusionsThis newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population.

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Section: Introductionmentioning

confidence: 99%

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

et al. 2012

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“…Additional genomic variance was subsequently described by Jager et al (36) in their RFLP analysis of 80 isolates. Differentiation of C. burnetii has also been achieved by sequence and/or PCR-RFLP analysis of icd (encoding isocitrate dehydrogenase) (5,46), com1 (encoding an outer membrane-associated immunoreactive protein) (34,62,83), and mucZ (known to confer a mucoid property to bacteria) (62). The most recent and extensive survey of C. burnetii genetic diversity was reported by Glazunova et al (24), who used multispacer sequence typing to place 173 C. burnetii isolates into 30 different genotypic groups.…”

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confidence: 99%

Genetic Diversity of the Q Fever Agent,Coxiella burnetii, Assessed by Microarray-Based Whole-Genome Comparisons

Beare¹,

Samuel

Howe³

et al. 2006

J Bacteriol

125

132

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Coxiella burnetii, a gram-negative obligate intracellular bacterium, causes human Q fever and is considered a potential agent of bioterrorism. Distinct genomic groups of C. burnetii are revealed by restriction fragmentlength polymorphisms (RFLP). Here we comprehensively define the genetic diversity of C. burnetii by hybridizing the genomes of 20 RFLP-grouped and four ungrouped isolates from disparate sources to a high-density custom Affymetrix GeneChip containing all open reading frames (ORFs) of the Nine Mile phase I (NMI) reference isolate. We confirmed the relatedness of RFLP-grouped isolates and showed that two ungrouped isolates represent distinct genomic groups. Isolates contained up to 20 genomic polymorphisms consisting of 1 to 18 ORFs each. These were mostly complete ORF deletions, although partial deletions, point mutations, and insertions were also identified. A total of 139 chromosomal and plasmid ORFs were polymorphic among all C. burnetii isolates, representing ca. 7% of the NMI coding capacity. Approximately 67% of all deleted ORFs were hypothetical, while 9% were annotated in NMI as nonfunctional (e.g., frameshifted). The remaining deleted ORFs were associated with diverse cellular functions. The only deletions associated with isogenic NMI variants of attenuated virulence were previously described large deletions containing genes involved in lipopolysaccharide (LPS) biosynthesis, suggesting that these polymorphisms alone are responsible for the lower virulence of these variants. Interestingly, a variant of the Australia QD isolate producing truncated LPS had no detectable deletions, indicating LPS truncation can occur via small genetic changes. Our results provide new insight into the genetic diversity and virulence potential of Coxiella species.

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“…In this regard, we conclude that we obtained robust and confirmed data to perform genomotyping with our microarray results. Several typing methods have been developed for the causative agent of Q fever [5], [7], [8], [10], [12]–[14]. Glazunova et al.…”

Section: Discussionmentioning

confidence: 99%

“…Restriction fragment length polymorphism (RFLP) analysis of genomic DNA (gDNA) [6]–[8] and sequence and/or PCR-RFLP analysis [9]–[12] of specific genes reveal genetic diversity between C. burnetii isolates. The most extensive survey of C. burnetii genetic diversity was reported by Glazunova et al [13], who used multi-spacer typing (MST) to genotype approximately 150 C. burnetii isolates.…”

Section: Introductionmentioning

confidence: 99%

Genomotyping of Coxiella burnetii Using Microarrays Reveals a Conserved Genomotype for Hard Tick Isolates

et al. 2011

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C. burnetii is a Gram-negative intracellular Y-proteobacteria that causes the zoonotic disease Q fever. Q fever can manifest as an acute or chronic illness. Different typing methods have been previously developed to classify C. burnetii isolates to explore its pathogenicity. Here, we report a comprehensive genomotyping method based on the presence or absence of genes using microarrays. The genomotyping method was then tested in 52 isolates obtained from different geographic areas, different hosts and patients with different clinical manifestations. The analysis revealed the presence of 10 genomotypes organized into 3 groups, with a topology congruent with that obtained through multi-spacer typing. We also found that only 4 genomotypes were specifically associated with acute Q fever, whereas all of the genomotypes could be associated to chronic human infection. Serendipitously, the genomotyping results revealed that all hard tick isolates, including the Nine Mile strain, belong to the same genomotype.

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Comparison of Japanese Isolates of Coxiella burnetii by PCR‐RFLP and Sequence Analysis

Cited by 12 publications

References 23 publications

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

Genetic Diversity of the Q Fever Agent,Coxiella burnetii, Assessed by Microarray-Based Whole-Genome Comparisons

Genomotyping of Coxiella burnetii Using Microarrays Reveals a Conserved Genomotype for Hard Tick Isolates

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