• 1.

    Flaherty BR, Barratt J, Lane M, Talundzic E, Bradbury RS , 2021. Sensitive universal detection of blood parasites by selective pathogen-DNA enrichment and deep amplicon sequencing. Microbiome 9: 119.

    • Search Google Scholar
    • Export Citation
  • 2.

    Flaherty BR, Talundzic E, Barratt J, Kines KJ, Olsen C, Lane M, Sheth M, Bradbury RS , 2018. Restriction enzyme digestion of host DNA enhances universal detection of parasitic pathogens in blood via targeted amplicon deep sequencing. Microbiome 6: 113.

    • Search Google Scholar
    • Export Citation
  • 3.

    Teal AE, Habura A, Ennis J, Keithly JS, Madison-Antenucci S , 2012. A new real-time PCR assay for improved detection of the parasite Babesia microti. J Clin Microbiol 50: 903908.

    • Search Google Scholar
    • Export Citation
  • 4.

    Shokoples SE, Ndao M, Kowalewska-Grochowska K, Yanow SK , 2009. Multiplexed real-time PCR assay for discrimination of Plasmodium species with improved sensitivity for mixed infections. J Clin Microbiol 47: 975980.

    • Search Google Scholar
    • Export Citation
  • 5.

    Rougemont M, Van Saanen M, Sahli R, Hinrikson HP, Bille J, Jaton K , 2004. Detection of four Plasmodium species in blood from humans by 18S rRNA gene subunit-based and species-specific real-time PCR assays. J Clin Microbiol 42: 56365643.

    • Search Google Scholar
    • Export Citation
  • 6.

    Fuehrer HP, Noedl H , 2014. Recent advances in detection of Plasmodium ovale: implications of separation into the two species Plasmodium ovale wallikeri and Plasmodium ovale curtisi. J Clin Microbiol 52: 387391.

    • Search Google Scholar
    • Export Citation
  • 7.

    Piron M, Fisa R, Casamitjana N, López-Chejade P, Puig L, Vergés M, Gascón J, Prat JGI, Portús M, Sauleda S , 2007. Development of a real-time PCR assay for Trypanosoma cruzi detection in blood samples. Acta Trop 103: 195200.

    • Search Google Scholar
    • Export Citation
  • 8.

    Ferreira LF, Britto C, Cardoso MA, Fernandes O, Reinhard K, Araujo A , 2000. Paleoparasitology of Chagas disease revealed by infected tissues from Chilean mummies. Acta Trop 75: 7984.

    • Search Google Scholar
    • Export Citation
  • 9.

    Galvao LM, Chiari E, Macedo AM, Luquetti AO, Silva SA, Andrade AL , 2003. PCR assay for monitoring Trypanosoma cruzi parasitemia in childhood after specific chemotherapy. J Clin Microbiol 41: 50665070.

    • Search Google Scholar
    • Export Citation
  • 10.

    Becker S, Franco JR, Simarro PP, Stich A, Abel PM, Steverding D , 2004. Real-time PCR for detection of Trypanosoma brucei in human blood samples. Diagn Microbiol Infect Dis 50: 193199.

    • Search Google Scholar
    • Export Citation
  • 11.

    Tellevik MG, Muller KE, Lokken KR, Nerland AH , 2014. Detection of a broad range of Leishmania species and determination of parasite load of infected mouse by real-time PCR targeting the arginine permease gene AAP3. Acta Trop 137: 99104.

    • Search Google Scholar
    • Export Citation
  • 12.

    Graca GC, Volpini AC, Romero GA, Oliveira Neto MP, Hueb M, Porrozzi R, Côrtes Boité M, Cupolillo E , 2012. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Mem Inst Oswaldo Cruz 107: 664674.

    • Search Google Scholar
    • Export Citation
  • 13.

    de Almeida ME, Steurer FJ, Koru O, Herwaldt BL, Pieniazek NJ, da Silva AJ , 2011. Identification of Leishmania spp. by molecular amplification and DNA sequencing analysis of a fragment of rRNA internal transcribed spacer 2. J Clin Microbiol 49: 31433149.

    • Search Google Scholar
    • Export Citation
  • 14.

    Kletsova EA, Spitzer ED, Fries BC, Marcos LA , 2017. Babesiosis in long island: review of 62 cases focusing on treatment with azithromycin and atovaquone. Ann Clin Microbiol Antimicrob 16: 17.

    • Search Google Scholar
    • Export Citation
  • 15.

    White DJ, Talarico J, Chang HG, Birkhead GS, Heimberger T, Morse DL , 1998. Human babesiosis in New York State: review of 139 hospitalized cases and analysis of prognostic factors. Arch Intern Med 158: 21492154.

    • Search Google Scholar
    • Export Citation
  • 16.

    Wright ES, Vetsigian KH , 2016. Quality filtering of illumina index reads mitigates sample cross-talk. BMC Genomics 17: 17.

  • 17.

    van der Valk T, Vezzi F, Ormestad M, Dalen L, Guschanski K , 2020. Index hopping on the Illumina HiseqX platform and its consequences for ancient DNA studies. Mol Ecol Resour 20: 11711181.

    • Search Google Scholar
    • Export Citation
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 

 

Assessing an Adaptation of the Universal Parasite Diagnostic Assay for Bloodborne Parasites in a US State Public Health Laboratory

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  • 1 New York State Department of Health, Wadsworth Center Parasitology Laboratory, Albany, New York;
  • | 2 Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, Parasitic Diseases Branch, Atlanta, Georgia;
  • | 3 Oak Ridge Institute for Science and Education, Oak Ridge, Tennessee;
  • | 4 Synergy America Inc., Duluth, Georgia;
  • | 5 SUNY Downstate Medical Center, Brooklyn, New York

ABSTRACT.

For complex clinical cases where a parasitic infection is suspected, it can be difficult for clinicians to recommend an appropriate laboratory test. These tests are usually pathogen-specific and require a certain degree of suspicion for the precise etiology. A recently described assay, the universal parasite diagnostic (UPDx) can potentially provide a diagnosis of any parasite present in a specimen. Using primers that amplify DNA from all eukaryotes, UPDx differentiates several parasitic infections in blood by amplicon-based next-generation sequencing (NGS) of the 18S rDNA locus. As the state’s public health reference laboratory, the Parasitology Laboratory at the Wadsworth Center (Albany, NY) receives specimens from patients who have potentially encountered a wide variety of parasites. As such, the ability to differentiate several blood parasites using a single assay is of interest. We assessed UPDx for its ability to confirm parasitic infections for 20 specimens that were previously identified by real-time PCR (RT-PCR). This included specimens positive for Babesia microti, Trypanosoma cruzi, Leishmania tropica, various Plasmodium species, and specimens comprising mixed Plasmodium sp. infections. Results obtained using UPDx were largely concordant with the RT-PCR assays. A T. cruzi positive specimen was negative by UPDx and for two mixed Plasmodium sp. infections only one species was detected. The results obtained for other specimens were concordant. We conclude that UPDx shows promise for the detection of blood parasites in diagnostic laboratories. As NGS becomes cheaper, assays like UPDx will become increasingly amenable to use in clinical settings.

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Author Notes

Address correspondence to Susan Madison-Antenucci, NYSDOH Wadsworth Center, Parasitology Laboratory, 120 New Scotland Avenue, Room 3112, Albany, NY 12201. E-mail: s.antenucci@health.ny.gov

Disclaimer: The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention/Agency for Toxic Substances and Disease Registry.

Authors’ addresses: Brooke Clemons, Allen E. Teal, and Susan Madison-Antenucci, New York State Department of Health, Wadsworth Center Parasitology Laboratory, Albany, NY, E-mails: brooke.clemons@health.ny.gov, allen.teal@health.ny.gov, and s.antenucci@health.ny.gov. Joel Barratt, Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, Parasitic Diseases Branch, Atlanta, GA, and Oak Ridge Institute for Science and Education, Oak Ridge, TN, E-mail: nsk9@cdc.gov. Yvonne Qvarnstrom, Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, Parasitic Diseases Branch, Atlanta, GA, E-mail: bvp2@cdc.gov. Meredith Lane, Synergy America Inc., Duluth, GA, E-mail: oeq5@cdc.gov. Greicy Zayas, SUNY Downstate Medical Center, Brooklyn, NY, E-mail: gzdlsm@gmail.com.

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