{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T20:37:06Z","timestamp":1793824626210,"version":"4.6.1"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,4,20]],"date-time":"2021-04-20T00:00:00Z","timestamp":1618876800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,4,20]],"date-time":"2021-04-20T00:00:00Z","timestamp":1618876800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2017YFD0501305"],"award-info":[{"award-number":["2017YFD0501305"]}]},{"name":"National Key Research and Development Program of China","award":["2019YFC1605700"],"award-info":[{"award-number":["2019YFC1605700"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U1904203"],"award-info":[{"award-number":["U1904203"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Cryptosporidium parvum<\/jats:italic>\n                      is an enteric protozoan parasite with zoonotic importance and can cause cryptosporidiosis in humans as well as domestic and wild animals worldwide. The IId subtype family (SF) is one of the most prevalent subtypes of\n                      <jats:italic>C. parvum<\/jats:italic>\n                      . Some clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) protein systems have been developed to detect nucleic acid with high flexibility, sensitivity and specificity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      By integrating recombinase polymerase amplification and the\n                      <jats:underline>C<\/jats:underline>\n                      as12a\/crRNA\n                      <jats:italic>\n                        <jats:underline>trans<\/jats:underline>\n                      <\/jats:italic>\n                      -\n                      <jats:underline>c<\/jats:underline>\n                      leavage system (termed ReCTC), we established end-point diagnostics by observing fluorescence readouts with the naked eye under blue light and on-site diagnostics using a lateral flow strip (LFS) biosensor.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Our ReCTC-based diagnoses can detect as little as a single copy of a cloned\n                      <jats:italic>C. parvum<\/jats:italic>\n                      60-kDa glycoprotein (GP60) gene, 10 oocysts per gram (OPG), clinical fecal sample without tedious extraction of genomic DNA and have no cross-reactivity with other SFs of\n                      <jats:italic>C. parvum<\/jats:italic>\n                      or other common enteric parasitic protozoa.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      This study provided a new strategy for direct identification of the IId SF of\n                      <jats:italic>C. parvum<\/jats:italic>\n                      free of highly trained operators and expensive special equipment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-021-04709-2","type":"journal-article","created":{"date-parts":[[2021,4,20]],"date-time":"2021-04-20T04:31:56Z","timestamp":1618893116000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":73,"title":["CRISPR\/Cas12a-based on-site diagnostics of Cryptosporidium parvum IId-subtype-family from human and cattle fecal samples"],"prefix":"10.1186","volume":"14","author":[{"given":"Fuchang","family":"Yu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaihui","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yilin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongfang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaohui","family":"Cui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianying","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sumei","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoying","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5706-131X","authenticated-orcid":false,"given":"Longxian","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,4,20]]},"reference":[{"key":"4709_CR1","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1128\/CMR.17.1.72-97.2004","volume":"17","author":"L Xiao","year":"2004","unstructured":"Xiao L, Fayer R, Ryan U, Upton SJ. Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev. 2004;17:72\u201397.","journal-title":"Clin Microbiol Rev"},{"key":"4709_CR2","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1016\/j.ijpara.2015.01.009","volume":"45","author":"U Ryan","year":"2015","unstructured":"Ryan U, Hijjawi N. New developments in Cryptosporidium research. Int J Parasitol. 2015;45:367\u201373.","journal-title":"Int J Parasitol"},{"key":"4709_CR3","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1038\/503189a","volume":"503","author":"B Striepen","year":"2013","unstructured":"Striepen B. Parasitic infections: time to tackle cryptosporidiosis. Nature. 2013;503:189\u201391.","journal-title":"Nature"},{"key":"4709_CR4","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1016\/j.actatropica.2018.08.018","volume":"187","author":"RJ Wang","year":"2018","unstructured":"Wang RJ, Li JQ, Chen YC, Zhang LX, Xiao LH. Widespread occurrence of Cryptosporidium infections in patients with HIV\/AIDS: Epidemiology, clinical feature, diagnosis, and therapy. Acta Trop. 2018;187:257\u201363.","journal-title":"Acta Trop"},{"key":"4709_CR5","doi-asserted-by":"publisher","first-page":"e28","DOI":"10.1590\/s1678-9946201961028","volume":"61","author":"FS Cunha","year":"2019","unstructured":"Cunha FS, Peralta JM, Peralta RHS. New insights into the detection and molecular characterization of Cryptosporidium with emphasis in Brazilian studies: a review. Rev Inst Med Trop Sao Paulo. 2019;61:e28.","journal-title":"Rev Inst Med Trop Sao Paulo"},{"key":"4709_CR6","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1186\/s13071-020-04191-2","volume":"13","author":"N Holubov\u00e1","year":"2020","unstructured":"Holubov\u00e1 N, T\u016fmov\u00e1 L, Sak B, Hejzlarov\u00e1 A, Kone\u010dn\u00fd R, McEvoy J, et al. Description of Cryptosporidium ornithophilus n. sp. (Apicomplexa: Cryptosporidiidae) in farmed ostriches. Parasit Vectors. 2020;13:340.","journal-title":"Parasit Vectors."},{"key":"4709_CR7","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1016\/j.exppara.2009.03.018","volume":"124","author":"L Xiao","year":"2010","unstructured":"Xiao L. Molecular epidemiology of cryptosporidiosis: an update. Exp Parasitol. 2010;124:80\u20139.","journal-title":"Exp Parasitol"},{"key":"4709_CR8","doi-asserted-by":"publisher","first-page":"1823","DOI":"10.3389\/fmicb.2017.01823","volume":"8","author":"R Wang","year":"2017","unstructured":"Wang R, Zhao G, Gong Y, Zhang L. Advances and perspectives on the epidemiology of bovine Cryptosporidium in China in the past 30 years. Front Microbiol. 2017;8:1823.","journal-title":"Front Microbiol"},{"key":"4709_CR9","doi-asserted-by":"publisher","first-page":"4208","DOI":"10.1038\/srep04208","volume":"4","author":"R Wang","year":"2014","unstructured":"Wang R, Zhang L, Axen C, Bjorkman C, Jian F, Amer S, et al. Cryptosporidium parvum IId family: clonal population and dispersal from Western Asia to other geographical regions. Sci Rep. 2014;4:4208.","journal-title":"Sci Rep"},{"key":"4709_CR10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.actatropica.2017.10.023","volume":"184","author":"A Khan","year":"2018","unstructured":"Khan A, Shaik JS, Grigg ME. Genomics and molecular epidemiology of Cryptosporidium species. Acta Trop. 2018;184:1\u201314.","journal-title":"Acta Trop"},{"key":"4709_CR11","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1126\/science.1231143","volume":"339","author":"L Cong","year":"2013","unstructured":"Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Multiplex genome engineering using CRISPR\/Cas systems. Science. 2013;339:819\u201323.","journal-title":"Science"},{"key":"4709_CR12","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1126\/science.1225829","volume":"337","author":"M Jinek","year":"2012","unstructured":"Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012;337:816\u201321.","journal-title":"Science"},{"key":"4709_CR13","doi-asserted-by":"publisher","first-page":"280","DOI":"10.1038\/nature24049","volume":"550","author":"OO Abudayyeh","year":"2017","unstructured":"Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, et al. RNA targeting with CRISPR-Cas13. Nature. 2017;550:280\u20134.","journal-title":"Nature"},{"key":"4709_CR14","doi-asserted-by":"publisher","first-page":"436","DOI":"10.1126\/science.aar6245","volume":"360","author":"JS Chen","year":"2018","unstructured":"Chen JS, Ma E, Harrington LB, Da Costa M, Tian X, Palefsky JM, et al. CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 2018;360:436\u20139.","journal-title":"Science"},{"key":"4709_CR15","doi-asserted-by":"publisher","first-page":"839","DOI":"10.1126\/science.aav4294","volume":"362","author":"LB Harrington","year":"2018","unstructured":"Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP, et al. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Science. 2018;362:839\u201342.","journal-title":"Science"},{"key":"4709_CR16","doi-asserted-by":"publisher","first-page":"1814","DOI":"10.1016\/j.cell.2016.11.053","volume":"167","author":"H Yang","year":"2016","unstructured":"Yang H, Gao P, Rajashankar KR, Patel DJ. PAM-dependent target DNA recognition and cleavage by C2c1 CRISPR-Cas endonuclease. Cell. 2016;167:1814\u201328.","journal-title":"Cell"},{"key":"4709_CR17","doi-asserted-by":"publisher","first-page":"438","DOI":"10.1126\/science.aam9321","volume":"356","author":"JS Gootenberg","year":"2017","unstructured":"Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung J, et al. Nucleic acid detection with CRISPR-Cas13a\/C2c2. Science. 2017;356:438\u201342.","journal-title":"Science"},{"key":"4709_CR18","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1038\/s41421-018-0028-z","volume":"4","author":"S Li","year":"2018","unstructured":"Li S, Cheng Q, Wang J, Li X, Zhang Z, Gao S, et al. CRISPR-Cas12a-assisted nucleic acid detection. Cell Discov. 2018;4:20.","journal-title":"Cell Discov"},{"key":"4709_CR19","doi-asserted-by":"publisher","first-page":"112068","DOI":"10.1016\/j.bios.2020.112068","volume":"154","author":"Q He","year":"2020","unstructured":"He Q, Yu D, Bao M, Korensky G, Chen J, Shin M, et al. High-throughput and all-solution phase African Swine Fever Virus (ASFV) detection using CRISPR-Cas12a and fluorescence based point-of-care system. Biosens Bioelectron. 2020;154:112068.","journal-title":"Biosens Bioelectron"},{"key":"4709_CR20","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1038\/s41421-020-0151-5","volume":"6","author":"S Lu","year":"2020","unstructured":"Lu S, Li F, Chen Q, Wu J, Duan J, Lei X, et al. Rapid detection of African swine fever virus using Cas12a-based portable paper diagnostics. Cell Discov. 2020;6:18.","journal-title":"Cell Discov"},{"key":"4709_CR21","doi-asserted-by":"publisher","first-page":"870","DOI":"10.1038\/s41587-020-0513-4","volume":"38","author":"JP Broughton","year":"2020","unstructured":"Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, et al. CRISPR-Cas12-based detection of SARS-CoV-2. Nat Biotechnol. 2020;38:870\u20134.","journal-title":"Nat Biotechnol"},{"key":"4709_CR22","doi-asserted-by":"publisher","first-page":"12156","DOI":"10.1021\/acs.analchem.9b01526","volume":"91","author":"B Wang","year":"2019","unstructured":"Wang B, Wang R, Wang D, Wu J, Wang J, Liu H, et al. Cas12aVDet: A CRISPR\/Cas12a-based platform for rapid and visual nucleic acid detection. Anal Chem. 2019;91:12156\u201361.","journal-title":"Anal Chem"},{"key":"4709_CR23","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1007\/s10096-019-03757-y","volume":"39","author":"G Xiao","year":"2020","unstructured":"Xiao G, Zhang S, Liang Z, Li G, Fang M, Liu Y, et al. Identification of Mycobacterium abscessus species and subspecies using the Cas12a\/sgRNA-based nucleic acid detection platform. Eur J Clin Microbiol Infect Dis. 2020;39:551\u20138.","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"4709_CR24","doi-asserted-by":"publisher","first-page":"514","DOI":"10.1186\/s13071-018-3088-x","volume":"11","author":"Z Cui","year":"2018","unstructured":"Cui Z, Song D, Qi M, Zhang S, Wang R, Jian F, et al. Revisiting the infectivity and pathogenicity of Cryptosporidium avium provides new information on parasitic sites within the host. Parasit Vectors. 2018;11:514.","journal-title":"Parasit Vectors"},{"key":"4709_CR25","doi-asserted-by":"publisher","first-page":"120818","DOI":"10.1016\/j.talanta.2020.120818","volume":"214","author":"M Zhang","year":"2020","unstructured":"Zhang M, Liu C, Shi Y, Wu J, Wu J, Chen H. Selective endpoint visualized detection of Vibrio parahaemolyticus with CRISPR\/Cas12a assisted PCR using thermal cycler for on-site application. Talanta. 2020;214:120818.","journal-title":"Talanta"},{"key":"4709_CR26","doi-asserted-by":"publisher","first-page":"28S","DOI":"10.1111\/j.1550-7408.2001.tb00442.x","volume":"48","author":"MM Peng","year":"2001","unstructured":"Peng MM, Matos O, Gatei W, Das P, Stantic-Pavlinic M, Bern C, et al. A comparison of Cryptosporidium subgenotypes from several geographic regions. J Eukaryot Microbiol. 2001;48:28S-31S.","journal-title":"J Eukaryot Microbiol."},{"key":"4709_CR27","doi-asserted-by":"publisher","first-page":"863","DOI":"10.1038\/nbt.3609","volume":"34","author":"D Kim","year":"2016","unstructured":"Kim D, Kim J, Hur JK, Been KW, Yoon SH, Kim JS. Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016;34:863\u20138.","journal-title":"Nat Biotechnol"},{"key":"4709_CR28","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1038\/nbt.3620","volume":"34","author":"BP Kleinstiver","year":"2016","unstructured":"Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM, et al. Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol. 2016;34:869\u201374.","journal-title":"Nat Biotechnol"},{"key":"4709_CR29","doi-asserted-by":"publisher","first-page":"730","DOI":"10.1016\/j.tibtech.2018.12.005","volume":"37","author":"Y Li","year":"2019","unstructured":"Li Y, Li S, Wang J, Liu G. CRISPR\/Cas systems towards next-generation biosensing. Trends Biotechnol. 2019;37:730\u201343.","journal-title":"Trends Biotechnol"}],"container-title":["Parasites &amp; Vectors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-021-04709-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13071-021-04709-2\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-021-04709-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,8]],"date-time":"2021-11-08T13:06:59Z","timestamp":1636376819000},"score":1,"resource":{"primary":{"URL":"https:\/\/parasitesandvectors.biomedcentral.com\/articles\/10.1186\/s13071-021-04709-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,20]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["4709"],"URL":"https:\/\/doi.org\/10.1186\/s13071-021-04709-2","relation":{},"ISSN":["1756-3305"],"issn-type":[{"value":"1756-3305","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,20]]},"assertion":[{"value":"2 January 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 March 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 April 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All research procedures used in this work were approved by the Institutional Review Board of Henan Agricultural University (approval no. IRB-HENAU-20190820-02). The collection of cattle fecal samples conformed to the Chinese Laboratory Animal Administration Act of 1988. The use of positive human samples in this work complied with Declaration of Helsinki 1975, as revised in 2013.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"208"}}