Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting Burkholderia cepacia Complex in Non-Sterile Pharmaceutical Products

被引：10

作者：

Daddy Gaoh, Soumana ^{[1
]}

Kweon, Ohgew ^{[1
]}

Lee, Yong-Jin ^{[2
]}

LiPuma, John J. ^{[3
]}

Hussong, David ^{[4
]}

Marasa, Bernard ^{[5
]}

Ahn, Youngbeom ^{[1
]}

机构：

[1] US FDA, Natl Ctr Toxicol Res, Div Microbiol, Jefferson, AR 72079 USA

[2] Albany State Univ, Dept Nat Sci, Albany, GA 31707 USA

[3] Univ Michigan, Dept Pediat, Ann Arbor, MI 48109 USA

[4] Eagle Analyt Serv, Houston, TX 77099 USA

[5] US FDA, Ctr Drug Evaluat & Res, Off Pharmaceut Qual, Silver Spring, MD 20993 USA

来源：

PATHOGENS | 2021年 / 10卷 / 09期

关键词：

Burkholderia cepacia complex; loop-mediated isothermal amplification (LAMP); nuclease-free water; antiseptics; IDENTIFICATION; RECOVERY; LIQUID;

D O I：

10.3390/pathogens10091071

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Simple and rapid detection of Burkholderia cepacia complex (BCC) bacteria, a common cause of pharmaceutical product recalls, is essential for consumer safety. In this study, we developed and evaluated a ribB-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the detection of BCC in (i) nuclease-free water after 361 days, (ii) 10 mu g/mL chlorhexidine gluconate (CHX) solutions, and (iii) 50 mu g/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/mu L for Burkholderia cenocepacia 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 ribB-based LAMP assay for the rapid and sensitive detection of BCC in pharmaceutical manufacturing.

引用

页数：16

共 38 条

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