{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T03:35:12Z","timestamp":1790048112495,"version":"4.0.1"},"reference-count":42,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,27]],"date-time":"2021-10-27T00:00:00Z","timestamp":1635292800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004606","name":"Chang Gung Memorial Hospital","doi-asserted-by":"publisher","award":["CMRPG3L0621"],"award-info":[{"award-number":["CMRPG3L0621"]}],"id":[{"id":"10.13039\/501100004606","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Group B Streptococcus (GBS) is an important pathogen of neonatal infections, and the clonal complex (CC)-17\/serotype III GBS strain has emerged as the dominant strain. The clinical manifestations of CC17\/III GBS sepsis may vary greatly but have not been well-investigated. A total of 103 CC17\/III GBS isolates that caused neonatal invasive diseases were studied using a new approach based on clustered regularly interspaced short palindromic repeats (CRISPR) loci and restriction fragment length polymorphism (RFLP) analyses. All spacers of CRISPR loci were sequenced and analyzed with the clinical presentations. After CRISPR-RFLP analyses, a total of 11 different patterns were observed among the 103 CRISPR-positive GBS isolates. GBS isolates with the same RFLP patterns were found to have highly comparable spacer contents. Comparative sequence analysis of the CRISPR1 spacer content revealed that it is highly diverse and consistent with the dynamics of this system. A total of 29 of 43 (67.4%) spacers displayed homology to reported phage and plasmid DNA sequences. In addition, all CC17\/III GBS isolates could be categorized into three subgroups based on the CRISPR-RFLP patterns and eBURST analysis. The CC17\/III GBS isolates with a specific CRISPR-RFLP pattern were more significantly associated with occurrences of severe sepsis (57.1% vs. 29.3%, p = 0.012) and meningitis (50.0% vs. 20.8%, p = 0.009) than GBS isolates with RFLP lengths between 1000 and 1300 bp. Whole-genome sequencing was also performed to verify the differences between CC17\/III GBS isolates with different CRISPR-RFLP patterns. We concluded that the CRISPR-RFLP analysis is potentially applicable to categorizing CC17\/III GBS isolates, and a specific CRISPR-RFLP pattern could be used as a new biomarker to predict meningitis and illness severity after further verification.<\/jats:p>","DOI":"10.3390\/ijms222111626","type":"journal-article","created":{"date-parts":[[2021,10,27]],"date-time":"2021-10-27T22:00:23Z","timestamp":1635372023000},"page":"11626","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Clustered Regularly Interspaced Short Palindromic Repeat Analysis of Clonal Complex 17 Serotype III Group B Streptococcus Strains Causing Neonatal Invasive Diseases"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0798-5378","authenticated-orcid":false,"given":"Jen-Fu","family":"Hsu","sequence":"first","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jang-Jih","family":"Lu","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"},{"name":"Department of Laboratory Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan"},{"name":"Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chih","family":"Lin","sequence":"additional","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shih-Ming","family":"Chu","sequence":"additional","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5858-6214","authenticated-orcid":false,"given":"Lee-Chung","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei-Yin","family":"Lai","sequence":"additional","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hsuan-Rong","family":"Huang","sequence":"additional","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ming-Chou","family":"Chiang","sequence":"additional","affiliation":[{"name":"Division of Pediatric Neonatology, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan"},{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0343-2370","authenticated-orcid":false,"given":"Ming-Horng","family":"Tsai","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan"},{"name":"Division of Neonatology and Pediatric Hematology\/Oncology, Department of Pediatrics, Chang Gung Memorial Hospital, Yunlin 638, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1016\/j.cmi.2015.05.039","article-title":"Group B streptococcus neonatal invasive infections, France 2007\u20132012","volume":"21","author":"Joubrel","year":"2015","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Lin, C., Chu, S.M., Wang, H.C., Yang, P.H., Huang, H.R., Chiang, M.C., Fu, R.H., Tsai, M.H., and Hsu, J.F. (2021). Complicated Streptococcus agalactiae sepsis with\/without meningitis in young infants and newborns: The clinical and molecular characteristics and outcomes. Microorganisms, 9.","DOI":"10.3390\/microorganisms9102094"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e791","DOI":"10.1093\/cid\/ciaa1485","article-title":"Group B Streptococcal disease in England (1998-2017): A population based observational study","volume":"72","author":"Kadambari","year":"2021","journal-title":"Clin. Infect Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1097\/INF.0000000000002269","article-title":"Neonatal group B Streptococcus infection in a tertiary care hospital in Saudi Arabia: A 13-year experience","volume":"38","author":"Madani","year":"2019","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"673.e9","DOI":"10.1016\/j.cmi.2017.02.029","article-title":"Group B streptococcal colonization and transmission dynamics in pregnant women and their newborns in Nigeria: Implications for prevention strategies","volume":"23","author":"Medugu","year":"2017","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Hsu, J.F., Tsai, M.H., Lin, L.C., Chu, S.M., Lai, M.Y., Huang, H.R., Chiang, M.C., Yang, P.H., and Lu, J.J. (2021). Genomic characterization of serotype III\/ST-17 Group B Streptococcus strains with antimicrobial resistance using whole genome sequencing. Biomedicines, 9.","DOI":"10.3390\/biomedicines9101477"},{"key":"ref_7","first-page":"374","article-title":"Antibiotic prevention for maternal group B Streptococcal colonization on neonatal GBS-related adverse outcomes: A meta-analysis","volume":"8","author":"Li","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1542\/peds.2013-1866","article-title":"2010 perinatal GBS prevention guideline and resource utilization","volume":"133","author":"Mukhopadhyay","year":"2014","journal-title":"Pediatrics"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1097\/INF.0000000000001414","article-title":"Neonatal group B streptococcus infections: Prevention strategies, clinical and microbiologic characteristics in 7 years of surveillance","volume":"36","author":"Creti","year":"2017","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1128\/JCM.00355-16","article-title":"Epidemiology of invasive group B streptococcal disease in Alberta, Canada, from 2003 to 2013","volume":"54","author":"Alhhazmi","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1016\/j.vaccine.2018.02.109","article-title":"Molecular characteristics of Streptococcus agalactiae in a mother-baby prospective cohort study: Implication for vaccine development and insights into vertical transmission","volume":"36","author":"Li","year":"2018","journal-title":"Vaccine"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Guan, X., Mu, X., Ji, W., Yuan, C., He, P., Zhang, L., Huang, Y., Li, J., Chen, J., and Zhong, H. (2018). Epidemiology of invasive group B streptococcal disease in infants from urban area of South China, 2011\u20132014. BMC Infect. Dis., 18.","DOI":"10.1186\/s12879-017-2811-0"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2695","DOI":"10.1128\/JCM.01183-16","article-title":"Molecular characteristics of Group B Streptococci isolated from adults with invasive infections in Japan","volume":"54","author":"Morozumi","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Li, C., Sapugahawatte, D., Yang, Y., Wong, K., Lo, N., and Ip, M. (2020). Multidrug-Resistant Streptococcus agalactiae Strains Found in Human and Fish with High Penicillin and Cefotaxime Non-Susceptibilities. Microorganisms, 8.","DOI":"10.3390\/microorganisms8071055"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"118525","DOI":"10.1016\/j.lfs.2020.118525","article-title":"CRISPR\/Cas9: A powerful genome editing technique for the treatment of cancer cells with present challenges and future directions","volume":"263","author":"Hazafa","year":"2020","journal-title":"Life Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"R40","DOI":"10.1093\/hmg\/ddu125","article-title":"CRISPR\/Cas9 for genome editing: Progress, implications and challenges","volume":"23","author":"Zhang","year":"2014","journal-title":"Hum. Mol. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5186","DOI":"10.1111\/febs.13110","article-title":"Genome modification by CRISPR\/Cas9","volume":"281","author":"Ma","year":"2014","journal-title":"FEBS J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"R79","DOI":"10.1093\/hmg\/ddy120","article-title":"Development and application of CRISPR\/Cas9 technologies in genomic editing","volume":"27","author":"Zhang","year":"2018","journal-title":"Hum. Mol. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Cho, S., Shin, J., and Cho, B.-K. (2018). Applications of CRISPR\/Cas System to Bacterial Metabolic Engineering. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19041089"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1930","DOI":"10.3389\/fmicb.2019.01930","article-title":"Cas9 contributes to group B Streptococcal colonization and diseases","volume":"10","author":"Spencer","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e01949-17","DOI":"10.1128\/JCM.01949-17","article-title":"Group B Streptococcus vaginal carriage in pregnant women as deciphered by clustered regularly interspaced short palindromic repeats analysis","volume":"56","author":"Beauruelle","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.3389\/fmicb.2017.01981","article-title":"CRISPR: A Useful Genetic Feature to Follow Vaginal Carriage of Group B Streptococcus","volume":"8","author":"Beauruelle","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1111\/j.1365-2958.2012.08172.x","article-title":"The highly dynamic crispr1 system of streptococcus agalactiae controls the diversity of its mobilome","volume":"85","author":"Sauvage","year":"2012","journal-title":"Mol. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Lier, C., Baticle, E., Horvath, P., Haguenoer, E., Valentin, A.-S., Glaser, P., Mereghetti, L., and Lanotte, P. (2015). Analysis of the type II-A CRISPR-Cas system of Streptococcus agalactiae reveals distinctive features according to genetic lineages. Front. Genet., 6.","DOI":"10.3389\/fgene.2015.00214"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/s41579-018-0071-7","article-title":"Molecular mechanisms of CRISPR\u2013Cas spacer acquisition","volume":"17","author":"McGinn","year":"2018","journal-title":"Nat. Rev. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"166","DOI":"10.3389\/fcimb.2019.00166","article-title":"phiD12-Like Livestock-Associated Prophages Are Associated with Novel Subpopulations of Streptococcus agalactiae Infecting Neonates","volume":"9","author":"Renard","year":"2019","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6682","DOI":"10.1016\/j.vaccine.2020.08.052","article-title":"Systematic review of Group B Streptococcal capsular types, sequence types and surface proteins as potential vaccine candidates","volume":"38","author":"Paul","year":"2020","journal-title":"Vaccine"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"129.e1","DOI":"10.1016\/j.cmi.2020.09.037","article-title":"Invasive group B Streptococcus infections in non-pregnant adults: A retrospective study, France, 2007\u20132019","volume":"27","author":"Vuillemin","year":"2020","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e01045-13","DOI":"10.1128\/mBio.01045-13","article-title":"Abundant and Diverse Clustered Regularly Interspaced Short Palindromic Repeat Spacers in Clostridium difficile Strains and Prophages Target Multiple Phage Types within This Pathogen","volume":"5","author":"Hargreaves","year":"2014","journal-title":"mBio"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1447","DOI":"10.3389\/fmicb.2019.01447","article-title":"Genomic Insights Into the Distribution and Evolution of Group B Streptococcus","volume":"10","author":"Chen","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"499","DOI":"10.3389\/fmicb.2017.00499","article-title":"Streptococcus agalactiae Causing Neonatal Infections in Portugal (2005\u20132015): Diversification and Emergence of a CC17\/PI-2b Multidrug Resistant Sublineage","volume":"8","author":"Martins","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.ijmm.2018.10.005","article-title":"Molecular epidemiology of invasive and non-invasive group B Streptococcus circulating in Serbia","volume":"309","author":"Gajic","year":"2018","journal-title":"Int. J. Med Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1089\/crispr.2020.0145","article-title":"Functional Study of the Type II-A CRISPR-Cas System of Streptococcus agalactiae Hypervirulent Strains","volume":"4","author":"Pastuszka","year":"2021","journal-title":"CRISPR J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1139\/cjm-2020-0212","article-title":"A short overview of the CRISPR-Cas adaptation stage","volume":"67","author":"Mosterd","year":"2021","journal-title":"Can. J. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e00728-20","DOI":"10.1128\/mBio.00728-20","article-title":"Pan-GWAS of Streptococcus agalactiae highlights lineage-specific genes associated with virulence and Niche adaptation","volume":"11","author":"Gori","year":"2020","journal-title":"mBio"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.3389\/fmicb.2016.01265","article-title":"Genomic analysis reveals multi-drug resistance clusters in group B Streptococcus CC17 hypervirulent isolates causing neonatal invasive disease in southern mainland China","volume":"7","author":"Campisi","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"264","DOI":"10.3389\/fmicb.2019.00264","article-title":"Molecular Characteristics and Antimicrobial Resistance of Group B Streptococcus Strains Causing Invasive Disease in Neonates and Adults","volume":"10","author":"Tsai","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Kao, Y., Tsai, M.H., Lai, M.Y., Chu, S.M., Huang, H.R., Chiang, M.C., Fu, R.H., Lu, J.J., and Hsu, J.F. (2019). Emerging serotype III sequence type 17 group B streptococcus invasive infection in infants: The clinical characteristics and impacts on outcomes. BMC Infect. Dis., 19.","DOI":"10.1186\/s12879-019-4177-y"},{"key":"ref_39","first-page":"379","article-title":"Changing epidemiology of group B streptococcal infections among adults in Iceland: 1975\u20132014","volume":"22","author":"Martins","year":"2015","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_40","unstructured":"Clinical and Laboratory Standards Institute (2014). Performance Standards for Antimicrobial Susceptibility Testing, Twenty-Fourth Informational Supplement, M100-S24, SLCI."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"574.e7","DOI":"10.1016\/j.cmi.2017.02.021","article-title":"Short-read whole genome sequencing for determination of antimicrobial resistance mechanisms and capsular serotypes of current invasive Streptococcus agalactiae recovered in the USA","volume":"23","author":"Metcalf","year":"2017","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"762","DOI":"10.3201\/eid2005.130953","article-title":"Streptococcus mitisStrains Causing Severe Clinical Disease in Cancer Patients","volume":"20","author":"Shelburne","year":"2014","journal-title":"Emerg. Infect. Dis."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11626\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:21:34Z","timestamp":1760167294000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11626"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,27]]},"references-count":42,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["ijms222111626"],"URL":"https:\/\/doi.org\/10.3390\/ijms222111626","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,27]]}}}