Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method

Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to estab...

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Main Authors: Chao Li, Gongyu Fu, Yaoqiang Shi, A-Mei Zhang, Xueshan Xia, Yue Fang, Xiaoqin Mao, Jie Jiang, Yuzhu Song, Guangying Yang
Format: Article
Language:English
Published: Associação Brasileira de Divulgação Científica 2019-03-01
Series:Brazilian Journal of Medical and Biological Research
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000300609&lng=en&tlng=en
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spelling doaj-2465c01b760447ec89384c8cca2083512020-11-25T01:36:59ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research1414-431X2019-03-0152310.1590/1414-431x20198186S0100-879X2019000300609Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification methodChao LiGongyu FuYaoqiang ShiA-Mei ZhangXueshan XiaYue FangXiaoqin MaoJie JiangYuzhu SongGuangying YangKlebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse blood with K. pneumoniae were used for sensitivity analysis. The detection limit of the LAMP was 1 bacterium/reaction. The results showed that the LAMP targeted to ureR_1 is a fast, specific, sensitive, inexpensive, and suitable method for the detection of K. pneumoniae.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000300609&lng=en&tlng=enLoop-mediated isothermal amplificationPolymerase chain reactionKlebsiella pneumoniaeNovel specific geneSpecificSensitive method
collection DOAJ
language English
format Article
sources DOAJ
author Chao Li
Gongyu Fu
Yaoqiang Shi
A-Mei Zhang
Xueshan Xia
Yue Fang
Xiaoqin Mao
Jie Jiang
Yuzhu Song
Guangying Yang
spellingShingle Chao Li
Gongyu Fu
Yaoqiang Shi
A-Mei Zhang
Xueshan Xia
Yue Fang
Xiaoqin Mao
Jie Jiang
Yuzhu Song
Guangying Yang
Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
Brazilian Journal of Medical and Biological Research
Loop-mediated isothermal amplification
Polymerase chain reaction
Klebsiella pneumoniae
Novel specific gene
Specific
Sensitive method
author_facet Chao Li
Gongyu Fu
Yaoqiang Shi
A-Mei Zhang
Xueshan Xia
Yue Fang
Xiaoqin Mao
Jie Jiang
Yuzhu Song
Guangying Yang
author_sort Chao Li
title Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
title_short Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
title_full Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
title_fullStr Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
title_full_unstemmed Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method
title_sort rapid, specific, and sensitive detection of the urer_1 gene in klebsiella pneumoniae by loop-mediated isothermal amplification method
publisher Associação Brasileira de Divulgação Científica
series Brazilian Journal of Medical and Biological Research
issn 1414-431X
publishDate 2019-03-01
description Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse blood with K. pneumoniae were used for sensitivity analysis. The detection limit of the LAMP was 1 bacterium/reaction. The results showed that the LAMP targeted to ureR_1 is a fast, specific, sensitive, inexpensive, and suitable method for the detection of K. pneumoniae.
topic Loop-mediated isothermal amplification
Polymerase chain reaction
Klebsiella pneumoniae
Novel specific gene
Specific
Sensitive method
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000300609&lng=en&tlng=en
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