Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products
Simple and rapid detection of <i>Burkholderia cepacia</i> complex (BCC) bacteria, a common cause of pharmaceutical product recalls, is essential for consumer safety. In this study, we developed and evaluated a <i>ribB</i>-based colorimetric loop-mediated isothermal amplificat...
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MDPI AG
2021-08-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/9/1071 |
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| author | Soumana Daddy Gaoh Ohgew Kweon Yong-Jin Lee John J. LiPuma David Hussong Bernard Marasa Youngbeom Ahn |
| author_facet | Soumana Daddy Gaoh Ohgew Kweon Yong-Jin Lee John J. LiPuma David Hussong Bernard Marasa Youngbeom Ahn |
| author_sort | Soumana Daddy Gaoh |
| collection | DOAJ |
| description | Simple and rapid detection of <i>Burkholderia cepacia</i> complex (BCC) bacteria, a common cause of pharmaceutical product recalls, is essential for consumer safety. In this study, we developed and evaluated a <i>ribB</i>-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the detection of BCC in (i) nuclease-free water after 361 days, (ii) 10 μg/mL chlorhexidine gluconate (CHX) solutions, and (iii) 50 μg/mL benzalkonium chloride (BZK) solutions after 184 days. The RibB 5 primer specifically detected 20 strains of BCC but not 36 non-BCC strains. The limit of detection of the LAMP assay was 1 pg/μL for <i>Burkholderia cenocepacia</i> strain J2315. Comparison of LAMP with a qPCR assay using 1440 test sets showed higher sensitivity: 60.6% in nuclease-free water and 42.4% in CHX solution with LAMP vs. 51.3% and 31.1%, respectively, with qPCR. These results demonstrate the potential of the <i>ribB</i>-based LAMP assay for the rapid and sensitive detection of BCC in pharmaceutical manufacturing. |
| first_indexed | 2024-03-10T07:20:37Z |
| format | Article |
| id | doaj.art-c6c03e5d9b3145998b0c3afc09cef5af |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T07:20:37Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-c6c03e5d9b3145998b0c3afc09cef5af2023-11-22T14:41:15ZengMDPI AGPathogens2076-08172021-08-01109107110.3390/pathogens10091071Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical ProductsSoumana Daddy Gaoh0Ohgew Kweon1Yong-Jin Lee2John J. LiPuma3David Hussong4Bernard Marasa5Youngbeom Ahn6Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USADivision of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USADepartment of Natural Sciences, Albany State University, Albany, GA 31707, USADepartment of Pediatrics, University of Michigan, Ann Arbor, MI 48109, USAEagle Analytical Services, Houston, TX 77099, USAOffice of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USASimple and rapid detection of <i>Burkholderia cepacia</i> complex (BCC) bacteria, a common cause of pharmaceutical product recalls, is essential for consumer safety. In this study, we developed and evaluated a <i>ribB</i>-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the detection of BCC in (i) nuclease-free water after 361 days, (ii) 10 μg/mL chlorhexidine gluconate (CHX) solutions, and (iii) 50 μg/mL benzalkonium chloride (BZK) solutions after 184 days. The RibB 5 primer specifically detected 20 strains of BCC but not 36 non-BCC strains. The limit of detection of the LAMP assay was 1 pg/μL for <i>Burkholderia cenocepacia</i> strain J2315. Comparison of LAMP with a qPCR assay using 1440 test sets showed higher sensitivity: 60.6% in nuclease-free water and 42.4% in CHX solution with LAMP vs. 51.3% and 31.1%, respectively, with qPCR. These results demonstrate the potential of the <i>ribB</i>-based LAMP assay for the rapid and sensitive detection of BCC in pharmaceutical manufacturing.https://www.mdpi.com/2076-0817/10/9/1071<i>Burkholderia cepacia</i> complexloop-mediated isothermal amplification (LAMP)nuclease-free waterantiseptics |
| spellingShingle | Soumana Daddy Gaoh Ohgew Kweon Yong-Jin Lee John J. LiPuma David Hussong Bernard Marasa Youngbeom Ahn Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products Pathogens <i>Burkholderia cepacia</i> complex loop-mediated isothermal amplification (LAMP) nuclease-free water antiseptics |
| title | Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products |
| title_full | Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products |
| title_fullStr | Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products |
| title_full_unstemmed | Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products |
| title_short | Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting <i>Burkholderia cepacia</i> Complex in Non-Sterile Pharmaceutical Products |
| title_sort | loop mediated isothermal amplification lamp assay for detecting i burkholderia cepacia i complex in non sterile pharmaceutical products |
| topic | <i>Burkholderia cepacia</i> complex loop-mediated isothermal amplification (LAMP) nuclease-free water antiseptics |
| url | https://www.mdpi.com/2076-0817/10/9/1071 |
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