Use of Southern Hybridization to Detect the Haemophilus Influenzae β-lactamase Gene

Hsiu-Chen Lin; Chih Chien Wang; Cheing Mei Yu; Mong Ling Chu

Use of Southern Hybridization to Detect the Haemophilus Influenzae β-lactamase Gene

貢獻的翻譯標題: 以南方墨點雜交法偵測流行性感冒嗜血桿菌的β-Lactamase基因

Hsiu-Chen Lin, Chih Chien Wang, Cheing Mei Yu, Mong Ling Chu

研究成果: 雜誌貢獻 › 文章 › 同行評審

摘要

會造成流行性感冒嗜血桿菌（Haemophilus influenzae）有ampicillin抗藥性的原因絕大多數是由於產生β-lactamase，而大部份的β-lactamase是屬於TEM-1型。但自1981年以來ROB-1型的β-lactamase 逐漸在流行性感冒嗜血桿菌及其他數種會造成動物感染疾病的菌株上發現。我們共收集了45株會產生β-lactamase的流行性感冒嗜血桿菌，用南方墨點雜交方法偵測其β-lactamase基因是屬於TEM-1型或ROB-1型。此方法是以TEM-1及ROB-1基因的DNA，以非放射性物質的digoxigenin標識當做探針，再與所有流行性感冒嗜血桿菌菌株的β-lactamase 之PCR產物做雜交反應。此次研究結果顯示，以南方墨點雜交方法偵測流行性感冒嗜血桿菌的β-lactamase可區分其屬於TEM-1型或ROB-1型。

貢獻的翻譯標題	以南方墨點雜交法偵測流行性感冒嗜血桿菌的β-Lactamase基因
原文	英語
頁（從 - 到）	16-20
頁數	5
期刊	中華民國感染症醫學會雜誌
卷	8
發行號	1
出版狀態	已發佈 - 6月 1997

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title = "Use of Southern Hybridization to Detect the Haemophilus Influenzae β-lactamase Gene",

abstract = "　The most common cause of ampicillin resistance in Haemophilus influenzae is due to the production of β-lactamase. In most strains, the β-lactamase produced is TEM-1. Since 1981, ROB-1 β-lactamase appears to be responsible for ampicillin resistance in H. influenzae and several animal pathogens. Forty-five ampicillin-resistant, β-lactamase-producing clinical isolates of Haemophilus influenzae were collected. We used Southern hybridization to determine whether they produced TEM-1 or ROB-1 β-lactamase. Plasmids were obtained from a ROB-1-producing strain of H. influenzae, and a plasmid (pBR322) known to encode TEM-1. Both β-lactamase genes were labeled with non-isotope digoxigenin as probes, and then hybridized on with PCR amplified β-lactamase gene product from H. influenzae strains. The results revealed that β-lactamase-mediated ampicillin resistance type in H. influenzae could be detected by the Southern hybridization method.",

keywords = "Southern hybridization, β-lactamase, Haemophilus influenzae",

author = "Hsiu-Chen Lin and Wang, {Chih Chien} and Yu, {Cheing Mei} and Chu, {Mong Ling}",

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T1 - Use of Southern Hybridization to Detect the Haemophilus Influenzae β-lactamase Gene

AU - Lin, Hsiu-Chen

AU - Wang, Chih Chien

AU - Yu, Cheing Mei

AU - Chu, Mong Ling

PY - 1997/6

Y1 - 1997/6

N2 - 　The most common cause of ampicillin resistance in Haemophilus influenzae is due to the production of β-lactamase. In most strains, the β-lactamase produced is TEM-1. Since 1981, ROB-1 β-lactamase appears to be responsible for ampicillin resistance in H. influenzae and several animal pathogens. Forty-five ampicillin-resistant, β-lactamase-producing clinical isolates of Haemophilus influenzae were collected. We used Southern hybridization to determine whether they produced TEM-1 or ROB-1 β-lactamase. Plasmids were obtained from a ROB-1-producing strain of H. influenzae, and a plasmid (pBR322) known to encode TEM-1. Both β-lactamase genes were labeled with non-isotope digoxigenin as probes, and then hybridized on with PCR amplified β-lactamase gene product from H. influenzae strains. The results revealed that β-lactamase-mediated ampicillin resistance type in H. influenzae could be detected by the Southern hybridization method.

AB - 　The most common cause of ampicillin resistance in Haemophilus influenzae is due to the production of β-lactamase. In most strains, the β-lactamase produced is TEM-1. Since 1981, ROB-1 β-lactamase appears to be responsible for ampicillin resistance in H. influenzae and several animal pathogens. Forty-five ampicillin-resistant, β-lactamase-producing clinical isolates of Haemophilus influenzae were collected. We used Southern hybridization to determine whether they produced TEM-1 or ROB-1 β-lactamase. Plasmids were obtained from a ROB-1-producing strain of H. influenzae, and a plasmid (pBR322) known to encode TEM-1. Both β-lactamase genes were labeled with non-isotope digoxigenin as probes, and then hybridized on with PCR amplified β-lactamase gene product from H. influenzae strains. The results revealed that β-lactamase-mediated ampicillin resistance type in H. influenzae could be detected by the Southern hybridization method.

KW - Southern hybridization

KW - β-lactamase

KW - Haemophilus influenzae

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Use of Southern Hybridization to Detect the Haemophilus Influenzae β-lactamase Gene

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