{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:17:05Z","timestamp":1760239025611,"version":"build-2065373602"},"reference-count":38,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,9,28]],"date-time":"2020-09-28T00:00:00Z","timestamp":1601251200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Apicomplexan parasites of the genus Babesia cause babesiosis in humans and animals worldwide. Human babesiosis is a predominantly zoonotic disease transmitted by hard ticks that is of increasing health concern in the USA and many other countries. Microscopic examination of stained blood smears, detection of serum antibodies by immunoassays and identification of parasite nucleic acid in blood by qPCR and fluorescence in situ hybridization (FISH) are some methods available for diagnosing babesiosis. This study investigated the use of a Babesia genus-specific FISH test for detecting Babesia parasites in blood smears and immunofluorescence assay (IFA) for detecting serum antibodies to Babesia duncani and Babesia microti, two common species that cause human babesiosis in the USA. The findings with clinical samples originating from USA, Australia, Europe and elsewhere demonstrate that the parallel use of Babesia genus-specific FISH and IFA tests for B. duncani and B. microti provides more useful diagnostic information in babesiosis and that B. duncani infections are more widespread globally than presently recognized.<\/jats:p>","DOI":"10.3390\/diagnostics10100761","type":"journal-article","created":{"date-parts":[[2020,9,28]],"date-time":"2020-09-28T08:02:58Z","timestamp":1601280178000},"page":"761","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Combined Immunofluorescence (IFA) and Fluorescence In Situ Hybridization (FISH) Assays for Diagnosing Babesiosis in Patients from the USA, Europe and Australia"],"prefix":"10.3390","volume":"10","author":[{"given":"Jyotsna S.","family":"Shah","sequence":"first","affiliation":[{"name":"ID-FISH Technology Inc., Milpitas, CA 95035, USA"},{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eddie","family":"Caoili","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie Fe","family":"Patton","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Snehal","family":"Tamhankar","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mu Mu","family":"Myint","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akhila","family":"Poruri","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olivia","family":"Mark","sequence":"additional","affiliation":[{"name":"IGenex Inc., 556 Gibraltar Drive, Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard I.","family":"Horowitz","sequence":"additional","affiliation":[{"name":"Hudson Valley Healing Arts Center, New York, NY 12538, USA"},{"name":"HHS Subcommittee on Babesia and Tick-Borne Pathogens, US Department of Health and Human Services, Washington, DC 20201, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alan D.","family":"Ashbaugh","sequence":"additional","affiliation":[{"name":"College of Medicine, University of Cincinnati, Cincinnati, OH 45221, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0246-7053","authenticated-orcid":false,"given":"Ranjan","family":"Ramasamy","sequence":"additional","affiliation":[{"name":"ID-FISH Technology Inc., Milpitas, CA 95035, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1016\/j.idc.2008.03.010","article-title":"Human babesiosis","volume":"22","author":"Vannier","year":"2008","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1007\/s40588-015-0025-z","article-title":"Human babesiosis: Pathogens, prevalence, diagnosis, and treatment","volume":"2","author":"Ord","year":"2015","journal-title":"Curr. Clin. Microbiol. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.ijpara.2018.11.007","article-title":"Human babesiosis","volume":"49","author":"Krause","year":"2019","journal-title":"Int. J. Parasitol."},{"key":"ref_4","unstructured":"Centers for Disease Control and Prevention (CDC) (2020). Surveillance for babesiosis\u2014United States, 2018 Annual Summary."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.ijpara.2018.07.002","article-title":"Evidence for transmission of the zoonotic apicomplexan parasite Babesia duncani by the tick Dermacentor albipictus","volume":"49","author":"Swei","year":"2019","journal-title":"Int. J. Parasitol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1869","DOI":"10.3201\/eid2211.151028","article-title":"Transmission of Babesia microti parasites by solid organ transplantation","volume":"22","author":"Brennan","year":"2016","journal-title":"Emerg. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.ijpara.2006.03.008","article-title":"Description of Babesia duncani n.sp. (Apicomplexa: Babesiidae) from humans and its differentiation from other piroplasms","volume":"36","author":"Conrad","year":"2006","journal-title":"Int. J. Parasitol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Scott, J.D., and Scott, C.M. (2018). Human babesiosis caused by Babesia duncani has widespread distribution across Canada. Healthcare, 6.","DOI":"10.3390\/healthcare6020049"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Shah, J.S., Mark, O., Caoili, E., Poruri, A., Horowitz, R.I., Ashbaugh, A.D., and Ramasamy, R. (2020). A Fluorescence in situ Hybridization (FISH) test for diagnosing babesiosis. Diagnostics, 10.","DOI":"10.3390\/diagnostics10060377"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.diagmicrobio.2015.03.010","article-title":"Comparison of a quantitative PCR assay with peripheral blood smear examination for detection and quantitation of Babesia microti infection in humans","volume":"82","author":"Wang","year":"2015","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1924","DOI":"10.1111\/trf.14642","article-title":"Superior real-time polymerase chain reaction detection of Babesia microti parasites in whole blood utilizing high-copy BMN antigens as amplification targets","volume":"58","author":"Grabias","year":"2018","journal-title":"Transfusion"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.4269\/ajtmh.16-0406","article-title":"Comparison of Babesia microti real-time polymerase chain reaction assays for confirmatory diagnosis of babesiosis","volume":"95","author":"Souza","year":"2016","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1916","DOI":"10.1111\/trf.14912","article-title":"Assessing the risk of Babesia to the United States blood supply using a risk-based decision-making approach: Report of AABB\u2019s ad hoc Babesia policy working group (original report)","volume":"58","author":"Ward","year":"2018","journal-title":"Transfusion"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1645\/17-155","article-title":"An improved PCR protocol for detection of Babesia duncani in wildlife and vector samples","volume":"104","author":"Kjemtrup","year":"2018","journal-title":"J. Parasitol."},{"key":"ref_15","first-page":"853","article-title":"Pathology of acute fatal babesiosis in hamsters experimentally infected with the WA-1 strain of Babesia","volume":"74","author":"Dao","year":"1996","journal-title":"Lab. Investig."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2037","DOI":"10.4049\/jimmunol.164.4.2037","article-title":"Selection of Babesia bovis-infected erythrocytes for adhesion to endothelial cells co-selects for altered variant erythrocyte surface antigen isoforms","volume":"164","author":"Allred","year":"2000","journal-title":"J. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/S0925-4439(97)00078-1","article-title":"Molecular basis for evasion of host immunity and pathogenesis in malaria","volume":"1406","author":"Ramasamy","year":"1998","journal-title":"Biochim. Biophys. Acta (BBA) Mol. Basis Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"14","DOI":"10.4269\/ajtmh.1978.27.14","article-title":"Babesia Microti infection in man: Evaluation of an indirect immunofluorescent antibody test","volume":"27","author":"Chisholm","year":"1978","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/infdis\/169.4.923","article-title":"Diagnosis of babesiosis: Evaluation of a serologic test for the detection of Babesia microti antibody","volume":"169","author":"Krause","year":"1994","journal-title":"J. Infect. Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1866","DOI":"10.1111\/trf.13618","article-title":"Serologic screening of United States blood donors for Babesia microti using an investigational enzyme immunoassay","volume":"56","author":"Levin","year":"2016","journal-title":"Transfusion"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Tayebwa, D.S., Beshbishy, A.M., Batiha, G.E.-S., Komugisha, M., Joseph, B., Vudriko, P., Yahia, R., Alkazmi, L., Hetta, H.F., and Yokoyama, N. (2020). Assessing the immunochromatographic test strip for serological detection of bovine babesiosis in Uganda. Microorganisms, 8.","DOI":"10.3390\/microorganisms8081110"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15585\/mmwr.ss6806a1","article-title":"Babesiosis surveillance\u2014United States, 2011-MMWR","volume":"68","author":"Gray","year":"2019","journal-title":"Surveill. Summ."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2014","DOI":"10.1128\/jcm.34.8.2014-2016.1996","article-title":"Efficacy of immunoglobulin M serodiagnostic test for rapid diagnosis of acute babesiosis","volume":"34","author":"Krause","year":"1996","journal-title":"J. Clin. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Bloch, E.M., Kumar, S., and Krause, P.J. (2019). Persistence of Babesia microti infection in humans. Pathogens, 8.","DOI":"10.3390\/pathogens8030102"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Harrison, T.E., M\u00f8rch, A.M., Felce, J.H., Sakoguchi, A., Reid, A.J., Arase, H., Dustin, M.L., and Higgins, M.K. (2020). Structural basis for RIFIN-mediated activation of LILRB1 in malaria. Nature, 1\u20137.","DOI":"10.1038\/s41586-020-2530-3"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"16079","DOI":"10.1038\/nmicrobiol.2016.79","article-title":"A global map of genetic diversity in Babesia microti reveals strong population structure and identifies variants associated with clinical relapse","volume":"1","author":"Lemieux","year":"2016","journal-title":"Nat. Microbiol."},{"key":"ref_27","unstructured":"(2020, July 31). Babesiosis and Tick-Borne Pathogens Subcommittee Report to the Tick-Borne Disease Working Group 2020, Available online: https:\/\/www.hhs.gov\/ash\/advisory-committees\/tickbornedisease\/reports\/babesiosis-subcomm-2020\/index.html."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"101","DOI":"10.2147\/IJGM.S193608","article-title":"Precision medicine: Retrospective chart review and data analysis of 200 patients on dapsone combination therapy for chronic Lyme disease\/post-treatment Lyme disease syndrome: Part 1","volume":"12","author":"Horowitz","year":"2019","journal-title":"Int. J. Gen. Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Liu, S., Du Cruz, I., Ramos, C.C., Taleon, P., Ramasamy, R., and Shah, J.S. (2018). Pilot study of immunoblots with recombinant Borrelia burgdorferi antigens for laboratory diagnosis of Lyme disease. Healthcare, 6.","DOI":"10.3390\/healthcare6030099"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Primus, S., Akoolo, L., Schlachter, S., Gedroic, K., Rojtman, A.D., and Parveen, N. (2018). Efficient detection of symptomatic and asymptomatic patient samples for Babesia microti and Borrelia burgdorferi infection by multiplex qPCR. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0196748"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Horowitz, R.I., and Freeman, P.R. (2018). Precision medicine: The role of the MSIDS model in defining, diagnosing, and treating chronic Lyme disease\/post treatment Lyme disease syndrome and other chronic illness: Part 2. Healthcare, 6.","DOI":"10.3390\/healthcare6040129"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1017\/S0031182006001387","article-title":"Evaluation of an indirect fluorescence immunoassay for the detection of serum antibodies against Babesia divergens in humans","volume":"134","author":"Duh","year":"2006","journal-title":"Parasitology"},{"key":"ref_33","first-page":"169","article-title":"Comparison of indirect fluorescent antibody (IFA) with complement fixation (CF) tests for diagnosis of Babesia spp infections in Colombian cattle","volume":"27","author":"Todorovic","year":"1976","journal-title":"Trop. Parasitol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1729","DOI":"10.1128\/CVI.00256-10","article-title":"Comparison of the Babesia duncani (WA1) IgG detection rates among clinical sera submitted to a reference laboratory for WA1 IgG testing and blood donor specimens from diverse geographic areas of the United States","volume":"17","author":"Prince","year":"2010","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"845","DOI":"10.2147\/IJGM.S27336","article-title":"Emerging incidence of Lyme borreliosis, babesiosis, bartonellosis, and granulocytic ehrlichiosis in Australia","volume":"4","author":"Mayne","year":"2011","journal-title":"Int. J. Gen. Med."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Young, K.M., Corrin, T., Wilhelm, B., Uhland, C., Greig, J., Mascarenhas, M., and Waddell, L.A. (2019). Zoonotic Babesia: A scoping review of the global evidence. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0226781"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"284","DOI":"10.7326\/0003-4819-119-4-199308150-00006","article-title":"Babesiosis in Washington state: A new species of Babesia?","volume":"119","author":"Quick","year":"1993","journal-title":"Ann. Intern. Med."},{"key":"ref_38","first-page":"507","article-title":"Comparative pathogenesis of human WA1 and Babesia microti isolates in a Syrian hamster model","volume":"46","author":"Wozniak","year":"1996","journal-title":"Lab. Anim. Sci."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/10\/761\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:14:23Z","timestamp":1760177663000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/10\/761"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,28]]},"references-count":38,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["diagnostics10100761"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics10100761","relation":{},"ISSN":["2075-4418"],"issn-type":[{"type":"electronic","value":"2075-4418"}],"subject":[],"published":{"date-parts":[[2020,9,28]]}}}