Details:

Alternative Title:
J Med Entomol
Personal Author:
GRAHAM, CHRISTINE B. ; BLACK, WILLIAM C. ; BOEGLER, KAREN A. ; MONTENIERI, JOHN A. ; HOLMES, JENNIFER L. ; ... More +
GRAHAM, CHRISTINE B. ; BLACK, WILLIAM C. ; BOEGLER, KAREN A. ; MONTENIERI, JOHN A. ; HOLMES, JENNIFER L. ; GAGE, KENNETH L. ; EISEN, REBECCA J. ; Less -
Description:
Programs that aim to control vector-borne zoonotic diseases require information on zoonotic hosts and on the feeding behavior of bridging vectors that are capable of transmitting pathogens from those hosts to humans. Here we describe an assay developed to identify bloodmeals in field-collected cat fleas (Ctenocephalides felis Bouché) to assess this species' potential role as a Yersinia pestis bridging vector in a plague-endemic region of Uganda. Our assay uses a single primer set and SYBR Green I-based real-time polymerase chain reaction to amplify a segment of the 12S mitochondrial ribosomal RNA gene for identification by sequencing. The assay capitalizes on the sensitivity of real-time polymerase chain reaction and the specificity of sequencing and can be used to differentiate vertebrate bloodmeals to the genus or species level without a priori knowledge of the host community. Because real-time assays that detect vertebrate DNA are highly sensitive to human DNA contamination, we analyzed detection in artificially fed and unfed fleas to establish a Ct cutoff that optimized specificity without completely sacrificing sensitivity. Using the established cutoff, our assay detected human, rat, and goat DNA in artificially fed C. felis up to 72 h postfeeding.
Subject:
[+]
Animals
Article
Bloodmeal Identification
Cats
Ctenocephalides
Cutoff
DNA
Flea
Fluorescent Dyes
Host Specificity
Humans
Organic Chemicals
Plague
Rats
Real-time PCR
Real-Time Polymerase Chain Reaction
Threshold Cycle
Yersinia Pestis

Source:
J Med Entomol. 49(6):1442-1452
Pubmed ID:
23270174
Pubmed Central ID:
PMC4642445
Document Type:
Journal Article ;
Funding:
CC999999/Intramural CDC HHS/United States ;
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:954c9e47337b7ecec9292204f731e46e67ca1c52f30c35b8d53eb3ca026786d8
File Type:

Supporting Files

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