Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction

Wilson, Kenneth H.; Blitchington, R B; Shah, Pratish; McDonald, Gregory A.; Gilmore, Robert D.; Mallavia, Louis P.

doi:10.1128/jcm.27.12.2692-2696.1989

Cited by 33 publications

(12 citation statements)

References 15 publications

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“…Since C. burnetii cannot be grown in axenic medium and has long been recovered from ticks, it has been classified in the Rickettsiales order, the Rickettsiaceae family, and the Rickettsiae tribe together with the genera Rickettsia and Rochalimaea (396). However, recent phylogenetic investigations, based mainly on 16S rRNA sequence analysis, have shown that the Coxiella genus belongs to the gamma subdivision of Proteobacteria (347,393,394,410), with the genera Legionella, Francisella, and Rickettsiella as its closest relatives (Fig. 1).…”

Section: Bacteriologymentioning

confidence: 99%

Q Fever

1999

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SUMMARY Q fever is a zoonosis with a worldwide distribution with the exception of New Zealand. The disease is caused by Coxiella burnetii, a strictly intracellular, gram-negative bacterium. Many species of mammals, birds, and ticks are reservoirs of C. burnetii in nature. C. burnetii infection is most often latent in animals, with persistent shedding of bacteria into the environment. However, in females intermittent high-level shedding occurs at the time of parturition, with millions of bacteria being released per gram of placenta. Humans are usually infected by contaminated aerosols from domestic animals, particularly after contact with parturient females and their birth products. Although often asymptomatic, Q fever may manifest in humans as an acute disease (mainly as a self-limited febrile illness, pneumonia, or hepatitis) or as a chronic disease (mainly endocarditis), especially in patients with previous valvulopathy and to a lesser extent in immunocompromised hosts and in pregnant women. Specific diagnosis of Q fever remains based upon serology. Immunoglobulin M (IgM) and IgG antiphase II antibodies are detected 2 to 3 weeks after infection with C. burnetii, whereas the presence of IgG antiphase I C. burnetii antibodies at titers of ≥1:800 by microimmunofluorescence is indicative of chronic Q fever. The tetracyclines are still considered the mainstay of antibiotic therapy of acute Q fever, whereas antibiotic combinations administered over prolonged periods are necessary to prevent relapses in Q fever endocarditis patients. Although the protective role of Q fever vaccination with whole-cell extracts has been established, the population which should be primarily vaccinated remains to be clearly identified. Vaccination should probably be considered in the population at high risk for Q fever endocarditis.

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Section: Bacteriologymentioning

confidence: 99%

Q Fever

1999

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“…Oligonucleotide probes directed at rRNA are therefore not sensitive enough to detect small populations of bacteria. To improve the sensitivity of rRNA probes, several investigators have taken the approach of using the polymerase chain reaction (PCR) primed with genusor speciesspecific oligonucleotides to amplify a ribosomal DNA (rDNA) gene fragment (2,3,5,6,16,19,28,39,43).…”

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confidence: 99%

Elimination of contaminating DNA within polymerase chain reaction reagents: implications for a general approach to detection of uncultured pathogens

et al. 1993

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Analysis based on comparisons of 16S rRNA sequences provides a rapid and reliable approach to identifying human pathogens. By directing oligonucleotide primers at sequences conserved throughout the eubacterial kingdom, bacterial 16S ribosomal DNA sequences of virtually any member of the eubacterial kingdom can be amplified by polymerase chain reaction and subsequently analyzed by sequence determination. Indeed, automated systems for broad-range amplification, sequencing, and data analysis are now feasible and may form the basis of the next generation of automated microbial identification systems. However, identification of pathogens by this strategy is hampered by the frequent contamination of reagents used for the amplification reaction, in particular Taq polymerase, with exogenous bacterial DNA. Here, we describe detailed investigations on the use of 8-methoxypsoralen and long-wave UV light to eliminate contaminating DNA in polymerase chain reaction reagents. The clinical utility of the developed procedure was demonstrated in a case of paucibacillary osteomyelitis, for which no specific bacterial agent had been cultured. rRNA sequences are an appealing target for detection, identification, and classification of bacteria (45). Compara-* Corresponding author.

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“…The PCR method has great potential for improving the ability to di agnose diseases caused by fastidious or slowly growing microorganisms (2) . It been employed for detection of rickettsiae , including R. rickettsii (24,26), R.…”

Section: Discussionmentioning

confidence: 99%