دورية أكاديمية
Polymerase chain reaction and loop-mediated isothermal amplification targeting lic13162, lic20239, and lipL32 genes for leptospirosis diagnosis.
| العنوان: | Polymerase chain reaction and loop-mediated isothermal amplification targeting lic13162, lic20239, and lipL32 genes for leptospirosis diagnosis. |
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| المؤلفون: | Pacce VD; Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, 96010-900, Brazil. violettapacce@gmail.com.; Laboratório de Biologia Molecular, Departamento de Engenharia de Bioprocessos E Biotecnologia, Universidade Federal Do Paraná, Curitiba, PR, 81280330, Brazil. violettapacce@gmail.com., Souza MN; Laboratório de Diagnóstico Molecular, Programa de Pós-Graduação Em Biologia Celular E Molecular Aplicada À Saúde, Universidade Luterana Do Brasil, Canoas, RS, Brazil., de Oliveira NR; Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, 96010-900, Brazil., Kremer FS; Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, 96010-900, Brazil., Jorge S; Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, 96010-900, Brazil., Ikuta N; Simbios Biotecnologia, Cachoeirinha, RS, Brazil., Lunge VR; Laboratório de Diagnóstico Molecular, Programa de Pós-Graduação Em Biologia Celular E Molecular Aplicada À Saúde, Universidade Luterana Do Brasil, Canoas, RS, Brazil.; Simbios Biotecnologia, Cachoeirinha, RS, Brazil., Dellagostin OA; Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, 96010-900, Brazil. |
| المصدر: | Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Braz J Microbiol] 2022 Jun; Vol. 53 (2), pp. 1029-1037. Date of Electronic Publication: 2022 Feb 05. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer International Publishing Country of Publication: Brazil NLM ID: 101095924 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1678-4405 (Electronic) Linking ISSN: 15178382 NLM ISO Abbreviation: Braz J Microbiol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2019- : Switzerland, AG : Springer International Publishing Original Publication: Rio de Janeiro, RJ, Brasil : Sociedade Brasileira de Microbiologia |
| مواضيع طبية MeSH: | Leptospira*/genetics , Leptospirosis*/diagnosis , Leptospirosis*/microbiology, Humans ; Molecular Diagnostic Techniques ; Nucleic Acid Amplification Techniques ; Real-Time Polymerase Chain Reaction/methods ; Sensitivity and Specificity |
| مستخلص: | Leptospirosis is a zoonotic disease caused by pathogenic species of Leptospira. Due to the similarity with clinical signs of other febrile diseases, early diagnosis remains challenging. Real-time PCR has been used for direct detection of Leptospira, but it requires thermocyclers and highly trained personnel. Loop-mediated isothermal amplification (LAMP) is a simple and rapid DNA-based assay. Therefore, here we have developed PCR and LAMP assays targeting two novel genes, lic13162 and lic20239, and also lipL32 gene to detect pathogenic Leptospira. Analytical and diagnostic performances were compared with bacterial isolates (including different Leptospira species and serovars) and clinical samples. The results demonstrated that PCR assays targeting lic13162 and lic20239 were successful to amplify Leptospira, but LAMP not. However, both PCR and LAMP targeting lipL32 could detect pathogenic Leptospira. LAMP lipL32 could be performed in 30 min with a detection limit of 156 cells/mL. Diagnostic performance of lipL32-LAMP presented 84.2% sensitivity and 93.2% specificity. In conclusion, lipL32 PCR and LAMP are effective methods to detect pathogenic Leptospira directly from clinical samples. (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.) |
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| فهرسة مساهمة: | Keywords: Accuracy; Diagnostic; Infectious diseases; LAMP; PCR; Point-of-care |
| SCR Protocol: | LAMP assay |
| تواريخ الأحداث: | Date Created: 20220206 Date Completed: 20220602 Latest Revision: 20230206 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC9151938 |
| DOI: | 10.1007/s42770-022-00698-1 |
| PMID: | 35124771 |
| قاعدة البيانات: | MEDLINE |
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Full Text Finder | تدمد: | 1678-4405 |
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| DOI: | 10.1007/s42770-022-00698-1 |