Differentiation of β-lactamase-negative ampicillin-resistant Haemophilus influenzae from other H. influenzae strains by a disc method

We established breakpoints for differentiating ampicillin (ABPC)-susceptible strains from resistant strains among Haemophilus influenzae isolates according to susceptibility to various beta-lactam antibiotics, using a disc method. Susceptibility testing of isolates for 13 beta-lactam agents was foll...

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Veröffentlicht in:Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy 2002-03, Vol.8 (1), p.50-58
Hauptverfasser: UBUKATA, Kimiko, CHIBA, Naoko, HASEGAWA, Keiko, SHIBASAKI, Yumi, SUNAKAWA, Keisuke, NONOYAMA, Masato, IWATA, Satoshi, KONNO, Masatoshi
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container_title Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy
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creator UBUKATA, Kimiko
CHIBA, Naoko
HASEGAWA, Keiko
SHIBASAKI, Yumi
SUNAKAWA, Keisuke
NONOYAMA, Masato
IWATA, Satoshi
KONNO, Masatoshi
description We established breakpoints for differentiating ampicillin (ABPC)-susceptible strains from resistant strains among Haemophilus influenzae isolates according to susceptibility to various beta-lactam antibiotics, using a disc method. Susceptibility testing of isolates for 13 beta-lactam agents was followed by analysis of the resistance genes, using a polymerase chain reaction (PCR) to identify the TEM-1 beta-lactamase gene ( bla) and the ftsI gene encoding penicillin-binding protein (PBP) 3, which affects beta-lactam minimum inhibitory concentrations (MICs). A total of 228 H. influenzae isolates were classified into 114 beta-lactamase-negative, ABPC-susceptible (BLNAS) strains; 29 beta-lactamase-negative, ABPC-resistant (BLNAR) strains; 53 low-BLNAR strains with a low degree of ABPC resistance; 27 TEM-1-producing strains (BLPAR); and 5 strains with ftsI gene mutations in addition to TEM-1 production (BLPACR) according to the PCR results. To identify resistant strains by disc-method susceptibility testing, the zone of inhibition was measured for ABPC (10 microg/disc), cefaclor (30 microg/disc), cefpodoxime (10 microg/disc), and cefdinir (5 microg/disc) discs. Strains were identified as BLNAS without resistant genes when the diameter was > or =27 mm for the ABPC disc and > or =21 mm for the cefaclor disc. Other strains were identified as BLNAR when the diameter was < or =22 mm for the cefpodoxime disc and < or =17 mm for the cefdinir disc. Remaining strains were identified as low-BLNAR. These criteria differentiated resistance types with high accuracy. A discrepancy was noted between genetic results and disc-testing breakpoints for differentiating resistant from susceptible H. influenzae. A disc-testing breakpoint for cefditoren (5 microg/disc) was proposed, with the susceptibility statistically defined as a diameter of > or =24 mm, which corresponds to the breakpoint (1 microg/ml) of the microdilution method recommended by the Japanese Society of Chemotherapy.
doi_str_mv 10.1007/s101560200006
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Susceptibility testing of isolates for 13 beta-lactam agents was followed by analysis of the resistance genes, using a polymerase chain reaction (PCR) to identify the TEM-1 beta-lactamase gene ( bla) and the ftsI gene encoding penicillin-binding protein (PBP) 3, which affects beta-lactam minimum inhibitory concentrations (MICs). A total of 228 H. influenzae isolates were classified into 114 beta-lactamase-negative, ABPC-susceptible (BLNAS) strains; 29 beta-lactamase-negative, ABPC-resistant (BLNAR) strains; 53 low-BLNAR strains with a low degree of ABPC resistance; 27 TEM-1-producing strains (BLPAR); and 5 strains with ftsI gene mutations in addition to TEM-1 production (BLPACR) according to the PCR results. To identify resistant strains by disc-method susceptibility testing, the zone of inhibition was measured for ABPC (10 microg/disc), cefaclor (30 microg/disc), cefpodoxime (10 microg/disc), and cefdinir (5 microg/disc) discs. Strains were identified as BLNAS without resistant genes when the diameter was &gt; or =27 mm for the ABPC disc and &gt; or =21 mm for the cefaclor disc. Other strains were identified as BLNAR when the diameter was &lt; or =22 mm for the cefpodoxime disc and &lt; or =17 mm for the cefdinir disc. Remaining strains were identified as low-BLNAR. These criteria differentiated resistance types with high accuracy. A discrepancy was noted between genetic results and disc-testing breakpoints for differentiating resistant from susceptible H. influenzae. A disc-testing breakpoint for cefditoren (5 microg/disc) was proposed, with the susceptibility statistically defined as a diameter of &gt; or =24 mm, which corresponds to the breakpoint (1 microg/ml) of the microdilution method recommended by the Japanese Society of Chemotherapy.</description><identifier>ISSN: 1341-321X</identifier><identifier>EISSN: 1437-7780</identifier><identifier>DOI: 10.1007/s101560200006</identifier><identifier>PMID: 11957120</identifier><language>eng</language><publisher>Tokyo: Springer</publisher><subject>Ampicillin Resistance ; Anti-Bacterial Agents - pharmacology ; Antibacterial agents ; Antibiotics. Antiinfectious agents. Antiparasitic agents ; beta-Lactamases - metabolism ; beta-Lactams ; Biological and medical sciences ; Haemophilus influenzae - drug effects ; Haemophilus influenzae - enzymology ; Humans ; Medical sciences ; Microbial Sensitivity Tests - methods ; Pharmacology. 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Strains were identified as BLNAS without resistant genes when the diameter was &gt; or =27 mm for the ABPC disc and &gt; or =21 mm for the cefaclor disc. Other strains were identified as BLNAR when the diameter was &lt; or =22 mm for the cefpodoxime disc and &lt; or =17 mm for the cefdinir disc. Remaining strains were identified as low-BLNAR. These criteria differentiated resistance types with high accuracy. A discrepancy was noted between genetic results and disc-testing breakpoints for differentiating resistant from susceptible H. influenzae. 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Susceptibility testing of isolates for 13 beta-lactam agents was followed by analysis of the resistance genes, using a polymerase chain reaction (PCR) to identify the TEM-1 beta-lactamase gene ( bla) and the ftsI gene encoding penicillin-binding protein (PBP) 3, which affects beta-lactam minimum inhibitory concentrations (MICs). A total of 228 H. influenzae isolates were classified into 114 beta-lactamase-negative, ABPC-susceptible (BLNAS) strains; 29 beta-lactamase-negative, ABPC-resistant (BLNAR) strains; 53 low-BLNAR strains with a low degree of ABPC resistance; 27 TEM-1-producing strains (BLPAR); and 5 strains with ftsI gene mutations in addition to TEM-1 production (BLPACR) according to the PCR results. To identify resistant strains by disc-method susceptibility testing, the zone of inhibition was measured for ABPC (10 microg/disc), cefaclor (30 microg/disc), cefpodoxime (10 microg/disc), and cefdinir (5 microg/disc) discs. Strains were identified as BLNAS without resistant genes when the diameter was &gt; or =27 mm for the ABPC disc and &gt; or =21 mm for the cefaclor disc. Other strains were identified as BLNAR when the diameter was &lt; or =22 mm for the cefpodoxime disc and &lt; or =17 mm for the cefdinir disc. Remaining strains were identified as low-BLNAR. These criteria differentiated resistance types with high accuracy. A discrepancy was noted between genetic results and disc-testing breakpoints for differentiating resistant from susceptible H. influenzae. A disc-testing breakpoint for cefditoren (5 microg/disc) was proposed, with the susceptibility statistically defined as a diameter of &gt; or =24 mm, which corresponds to the breakpoint (1 microg/ml) of the microdilution method recommended by the Japanese Society of Chemotherapy.</abstract><cop>Tokyo</cop><pub>Springer</pub><pmid>11957120</pmid><doi>10.1007/s101560200006</doi><tpages>9</tpages></addata></record>
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subjects Ampicillin Resistance
Anti-Bacterial Agents - pharmacology
Antibacterial agents
Antibiotics. Antiinfectious agents. Antiparasitic agents
beta-Lactamases - metabolism
beta-Lactams
Biological and medical sciences
Haemophilus influenzae - drug effects
Haemophilus influenzae - enzymology
Humans
Medical sciences
Microbial Sensitivity Tests - methods
Pharmacology. Drug treatments
title Differentiation of β-lactamase-negative ampicillin-resistant Haemophilus influenzae from other H. influenzae strains by a disc method
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