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Quantifying mosquito biting patterns on humans by DNA fingerprinting of bloodmeals.

E MichaelWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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K D RamaiahWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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S L HotiWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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G BarkerWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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M R PaulWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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J YuvarajWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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P K DasWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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B T GrenfellWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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D A BundyWellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, United Kingdom. e.michael@ic.ac.uk

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A major debate in infectious disease epidemiology concerns the relative importance of exposure and host factors, such as sex and acquired immunity, in determining observed age patterns of parasitic infection in endemic communities. Nonhomogeneous contact between hosts and vectors is also expected to increase the reproductive rate, and hence transmission, of mosquito-borne infections. Resolution of these questions for human parasitic diseases has been frustrated by the lack of a quantitative tool for quantifying the exposure rate of people in communities. Here, we show that the polymerase chain reaction (PCR) technique for amplifying and fingerprinting human DNA from mosquito bloodmeals can address this problem for mosquito-borne diseases. Analysis of parallel human and mosquito (resting Culex quinquefasciatus) samples from the same households in an urban endemic focus for bancroftian filariasis in South India demonstrates that a 9-locus radioactive short-tandem repeat system is able to identify the source of human DNA within the bloodmeals of nearly 80% of mosquitoes. The results show that a person's exposure rate, and hence the age and sex patterns of exposure to bites in an endemic community, can be successfully quantified by this method. Out of 276 bloodmeal PCR fingerprints, we also found that on average, 27% of the mosquitoes caught resting within individual households had fed on people outside the household. Additionally, 13% of mosquitoes biting within households contained blood from at least 2 people, with the rate of multiple feeding depending on the density of humans in the household. These complex vector feeding behaviors may partly account for the discrepancies in estimates of the infection rates of mosquito-borne diseases calculated parasitologically and entomologically, and they underline the potential of this tool for investigating the transmission dynamics of infection.

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