Evaluation of conventional and real-time PCR assays for detection and differentiation of Spotted Fever Group Rickettsia in dog blood

Kidd, Linda; Maggi, Ricardo G.; Diniz, Pedro; Hegarty, Barbara C.; Tucker, Melissa D.; Breitschwerdt, Edward B.

doi:10.1016/j.vetmic.2007.11.035

Cited by 98 publications

(83 citation statements)

References 21 publications

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“…A new step for the diagnosis of tick-borne rickettsioses has been achieved recently with the emergence of the use of quantitative real-time PCR (467)(468)(469). Genomic approaches have recently increased our knowledge of the genus Rickettsia, and massive amounts of genomic data have become available (1,470).…”

Section: New Approaches To Diagnosismentioning

confidence: 99%

Update on Tick-Borne Rickettsioses around the World: a Geographic Approach

et al. 2013

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SUMMARY Tick-borne rickettsioses are caused by obligate intracellular bacteria belonging to the spotted fever group of the genus Rickettsia . These zoonoses are among the oldest known vector-borne diseases. However, in the past 25 years, the scope and importance of the recognized tick-associated rickettsial pathogens have increased dramatically, making this complex of diseases an ideal paradigm for the understanding of emerging and reemerging infections. Several species of tick-borne rickettsiae that were considered nonpathogenic for decades are now associated with human infections, and novel Rickettsia species of undetermined pathogenicity continue to be detected in or isolated from ticks around the world. This remarkable expansion of information has been driven largely by the use of molecular techniques that have facilitated the identification of novel and previously recognized rickettsiae in ticks. New approaches, such as swabbing of eschars to obtain material to be tested by PCR, have emerged in recent years and have played a role in describing emerging tick-borne rickettsioses. Here, we present the current knowledge on tick-borne rickettsiae and rickettsioses using a geographic approach toward the epidemiology of these diseases.

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Section: New Approaches To Diagnosismentioning

confidence: 99%

Update on Tick-Borne Rickettsioses around the World: a Geographic Approach

et al. 2013

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“…and viruses (Bode et al 2004, Kocagoz et al 2005, Takahashi et al 2007, Tomaso et al 2007, Abril et al 2008, Kidd et al 2008. In recent years, fish disease diagnosticians have used this technique to identify and quantify bacterial, viral, and parasitic fish pathogens such as Aeromonas salmonicida, Flavobacterium columnare, Renibacterium salmoninarum, Henneguya ictaluri, largemouth bass virus, and recently Francisella piscicida in Norwegian cod (Balcázar et al 2007, Getchell et al 2007, Panangala et al 2007, Suzuki & Sakai 2007, Griffin et al 2008, Ottem et al 2008.…”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Soto¹,

Bowles²,

Fernandez³

et al. 2010

Dis. Aquat. Org.

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Members of the genus Francisella are small Gram-negative facultative intracellular bacteria that cause francisellosis in a wide variety of fish species worldwide. F. noatunensis subsp. orientalis has been recently described as a warm-water pathogen of tilapia Oreochromis spp. In this study, a quantitative real-time polymerase chain reaction (qPCR) TaqMan probe assay was developed to rapidly and accurately detect and quantify F. noatunensis subsp. orientalis from fish tissue. The target region of the assay was the F. tularensis iglC gene homologue previously found in F. noatunensis subsp. orientalis. Probe specificity was confirmed by the lack of signal and cross-reactivity with 12 common fish pathogens, 2 subspecies of F. tularensis, F. noatunensis subsp. noatunensis, and tilapia tissue. The range of linearity was determined to be 50 fg to 1.4 mg, and the lower limit of detection was 50 fg of DNA (equivalent to ~25 genome equivalents) per reaction. A similar sensitivity was observed with DNA extracted from a mixture of F. noatunensis subsp. orientalis and fish tissue. The assay was also able to detect and quantify F. noatunensis subsp. orientalis from the spleens of experimentally infected tilapia. No signal was observed in the control groups. In conclusion, we have developed a highly sensitive and specific assay that can be used for the specific identification and quantification of F. noatunensis subsp. orientalis. KEY WORDS: Francisella · Tilapia · qPCRResale or republication not permitted without written consent of the publisher

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“…DNA from B. henselae (similar to this sequence under GenBank accession number BX897699) was used as a positive control for rpoB PCR. Spotted fever group Rickettsia DNA was amplified by real-time PCR with Sybr green as a detection method targeting a fragment of the outer membrane protein A (ompA) gene as described previously (20). Rickettsia conorii DNA (similar to the sequence under GenBank accession number DQ518245) was used as a positive control for ompA PCR.…”

mentioning

confidence: 99%

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

Otranto¹,

Testini²,

Dantas‐Torres³

et al. 2010

J Clin Microbiol

100

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Canine vector-borne diseases (CVBDs) pose a diagnostic challenge, particularly when a dog is coinfected with more than one pathogen. The purpose of this study was to generate information about the diagnosis of CVBDs in young dogs following their first exposure to flea, tick, sand fly, louse, and mosquito vectors. From March 2008 to May 2009, 10 purpose-bred young naive beagle dogs and a cohort of 48 mixed-breed dogs living in an area to which CVBD is endemic in southern Italy were monitored using different diagnostic tests (cytology, serology, and PCR). Overall, PCR detected the highest number of dogs infected with Anaplasma platys, Babesia vogeli, and Ehrlichia canis, whereas seroconversion was a more sensitive indicator of exposure to Leishmania infantum. For A. platys infection, combining blood and buffy coat cytology in parallel enhanced the relative sensitivity (SE rel ) (87.3%). For B. vogeli, the best diagnostic combination was buffy coat cytology and serology used in parallel (SE rel , 67.5%), whereas serology and PCR used in parallel (SE rel , 100%) was the best combination for L. infantum. Overall, 12 (20.7%) dogs were coinfected; however, the percentage of new coinfections decreased from baseline (50%) to the first (33.3%) and second (16.6%) follow-up time points. Numbers of coinfections with A. platys and B. vogeli were significantly higher (P < 0.05) than coinfections with other pathogen combinations. The data generated in this study provide insights on the incidence of certain pathogens infecting young dogs in southern Italy, highlight important diagnostic testing limitations, and support the use of multiple diagnostic modalities when attempting to confirm a tick-borne infection in an individual dog or in a canine population.

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Evaluation of conventional and real-time PCR assays for detection and differentiation of Spotted Fever Group Rickettsia in dog blood

Cited by 98 publications

References 21 publications

Update on Tick-Borne Rickettsioses around the World: a Geographic Approach

Update on Tick-Borne Rickettsioses around the World: a Geographic Approach

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

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