Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

doi:10.1016/j.exppara.2014.12.003

Abstract

Babesia spp. are obligate protozoan parasites of red blood cells. Transmission to humans occurs through bites from infected ticks or blood transfusion. Infections with B. microti account for the majority of the reported cases of human babesiosis in the USA. A lower incidence is caused by the more recently described species B. duncani. The current gold standard for detection of Babesia is microscopic examination of blood smears. Recent PCR-based assays, including real-time PCR, have been developed for B. microti. On the other hand, molecular assays that detect and distinguish between B. microti and B. duncani infections are lacking. Closely related species of Babesia can be differentiated due to sequence variation within the internal transcribed spacer (ITS) regions of nuclear ribosomal RNAs. In the present study, we targeted the ITS regions of B. microti and B. duncani to develop sensitive and species-specific droplet digital PCR (ddPCR) assays. The assays were shown to discriminate B. microti from B. duncaniand resulted in limits of detection of ~10 gene copies. Moreover, ddPCR for these species were useful in DNA extracted from blood of experimentally infected hamsters, detecting infections of low parasitemia that were negative by microscopic examination. In summary, we have developed sensitive and specific quantitative ddPCR assays for the detection of B. microti and B. duncani in blood. Our methods could be used as sensitive approaches to monitor the progression of parasitemia in rodent models of infection as well as serve as suitable molecular tests in blood screening.

  1. I was just diagnosed this morning and don’t know how long I’ve had it other than a bullet rash I got about 2.5 weeks ago. My Babesia micro IgG Titer is 1-160H and the IgM Titer is 1-10 so that’s normal.

    I’m waiting for my physician to get back to me and was hoping someone, specifically a Dr., would be able to comment upon the newest and effective treatments.

    The only symptoms other than the rash have been:

    Slight pain and swelling in my middle knuckles on both hands (thought it was arthritis) I’m a very health 55 year old.

    Some bouts of extremely tiredness where I just had to lay down and nap for 2-3 hours. I’ve felt fine this week but am in the midst of a lot of work so I’ve been drinking a lot of coffee to combat any potential tiredness.

    Some mild headaches. This is not normal for me.

    I have a red, small dot rash on the lower part of my stomach but I think that is from humidity.

    FYI my 10 lb dog is also in the vets and they think she may have liptotenosis are we’re awaiting the blood test results. I shared with them vet that I have Babesia.

    Best and many thanks in advance for any assistance,

    Maureen D.

  2. david on said:

    I just got positive for Babesia FISH test

    Had to go all the way to brussels to get tested, original igenex test was negative when i irst tested it, on top of that Yersinia , psodonomas and some other things..

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