Detection of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti, with two different multiplex PCR assays

Hojgaard, Andrias; Lukacik, Gary; Piesman, Joseph

doi:10.1016/j.ttbdis.2013.12.001

Cited by 61 publications

(53 citation statements)

References 7 publications

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“…Recently, the use of real-time PCR has been described for detection of B. microti in ticks or clinical specimens from patients suspected of having babesiosis (Bloch et al, 2013;Chan et al, 2013;Hojgaard et al, 2014;Rollend et al, 2013;Teal et al, 2012). In these reports, the analytical sensitivity of real-time PCR assays was most often assessed by spiking plasmid DNA to negative blood samples, which may result in inaccurate results due to the potential for preferred amplification by PCR of small plasmid DNA fragments, compared with the same target sequence contained within the complete genome of a microorganism (Lin et al, 2011;Yun et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Utilization of a real-time PCR assay for diagnosis of Babesia microti infection in clinical practice

Wang

Wormser

Zhuge

et al. 2015

Ticks and Tick-borne Diseases

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Section: Introductionmentioning

confidence: 99%

Utilization of a real-time PCR assay for diagnosis of Babesia microti infection in clinical practice

Wang

Wormser

Zhuge

et al. 2015

Ticks and Tick-borne Diseases

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“…PCR and DNA purification controls included the I. scapularis actin target (Hojgaard et al, 2014) for tick samples and the rodent GAPDH target (Applied Biosystems ® TaqMan ® Rodent GAPDH ControlReagents kit; ThermoFisher) for mouse blood samples.…”

Section: Methodsmentioning

confidence: 99%

Transmission of the relapsing fever spirochete, Borrelia miyamotoi, by single transovarially-infected larval Ixodes scapularis ticks

Breuner

Hojgaard

Replogle

et al. 2018

Ticks and Tick-borne Diseases

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The relapsing fever spirochete, Borrelia miyamotoi, is increasingly recognized as a cause of human illness (hard tick-borne relapsing fever) in the United States. We previously demonstrated that single nymphs of the blacklegged tick, Ixodes scapularis, can transmit B. miyamotoi to experimental hosts. However, two recent epidemiological studies from the Northeastern United States indicate that human cases of hard tick-borne relapsing fever peak during late summer, after the spring peak for nymphal tick activity but coincident with the peak seasonal activity period of larval ticks in the Northeast. These epidemiological findings, together with evidence that B. miyamotoi can be passed from infected I. scapularis females to their offspring, suggest that bites by transovarially-infected larval ticks can be an important source of human infection. To demonstrate experimentally that transovarially-infected larval I. scapularis ticks can transmit B. miyamotoi, outbred Mus musculus CD1 mice were exposed to 1 or 2 potentially infected larvae. Individual fed larvae and mouse blood taken 10 d after larvae attached were tested for presence of B. miyamotoi DNA, and mice also were examined for seroreactivity to B. miyamotoi 8 wk after tick feeding. We documented B. miyamotoi DNA in blood from 13 (57%) of 23 mice exposed to a single transovarially-infected larva and in 5 (83%) of 6 mice exposed to two infected larvae feeding simultaneously. All 18 positive mice also demonstrated seroreactivity to B. miyamotoi. Of the 11 remaining mice without detectable B. miyamotoi DNA in their blood 10 d after infected larvae attached, 7 (64%) had evidence of spirochete exposure by serology 8 wk later. Because public health messaging for risk of exposure to Lyme disease spirochetes focuses on nymphal and female I. scapularis ticks, our finding that transovarially-infected larvae effectively transmit B. miyamotoi should lead to refined tick-bite prevention messages.

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“…Nucleic acids were isolated from ticks using DNeasy Blood and Tissue kit (Qiagen, Valencia, CA) and a Mini-Beadbeater (Biospec, Bartlesville, OK; Hojgaard et al 2014). To test for the presence of B. burgdorferi, a multiplex TaqMan PCR reaction was performed, targeting both B. burgdorferi and I. scapularis.…”

Section: Methodsmentioning

confidence: 99%

“…To test for the presence of B. burgdorferi, a multiplex TaqMan PCR reaction was performed, targeting both B. burgdorferi and I. scapularis. As a control for both the DNA purification and the PCR reaction, a set of primers and probe against the actin gene of I. scapularis was used (Hojgaard et al 2014). For detection of B. burgdorferi DNA, previously described primers and probes for the flagellar filament cap gene (fliD) were used (Dolan et al 2011).…”

Section: Methodsmentioning

confidence: 99%

Comparison of Tick Feeding Success and Vector Competence for Borrelia burgdorferi Among Immature Ixodes scapularis (Ixodida: Ixodidae) of Both Southern and Northern Clades

Goddard

Embers

Hojgaard

et al. 2015

Journal of Medical Entomology

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Northern and southern Ixodes scapularis Say populations differ greatly in density, host utilization, and especially questing behavior of the immatures. Haplotypes of I. scapularis in North America can be divided into two major clades-the All American Clade (haplotypes A through J) and the Southern Clade (M through O). This genetic variation may affect feeding success and vector competence. This study compared feeding success of larval I. scapularis measured by time-to-drop-off and subsequent transmissibility success of Borrelia burgdorferi to mice using ticks from Mississippi, Connecticut (both F haplotype), and Louisiana (haplotype O). Northern ticks (CT) fed to repletion much faster than MS and LA ticks: overall, 73.6% of CT ticks had dropped off mice at Day 3 compared to only 1.7% and 6.6% of ticks dropped off for MS and LA ticks at that same time point. As for vector competence, 4 of the 4 mice in each case (MS or CT) that had been fed on by infected nymphs tested positive for B. burgdorferi. In a second experiment, 5 of the 6 mice tested positive for B. burgdorferi after exposure to infected LA ticks as compared with 3 of the 4 mice exposed to infected CT ticks. These data demonstrate that there is no difference in northern and southern populations of I. scapularis in their ability to transmit B. burgdorferi, but the ability of the northern populations to feed rapidly on rodents exceeds that of southern populations.

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Detection of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti, with two different multiplex PCR assays

Cited by 61 publications

References 7 publications

Utilization of a real-time PCR assay for diagnosis of Babesia microti infection in clinical practice

Utilization of a real-time PCR assay for diagnosis of Babesia microti infection in clinical practice

Transmission of the relapsing fever spirochete, Borrelia miyamotoi, by single transovarially-infected larval Ixodes scapularis ticks

Comparison of Tick Feeding Success and Vector Competence for Borrelia burgdorferi Among Immature Ixodes scapularis (Ixodida: Ixodidae) of Both Southern and Northern Clades

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