Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients
The increasing incidence of azole resistance in Aspergillus fumigatus causing invasive aspergillosis (IA) in immunocompromised/hematological patients emphasizes the need to improve the detection of resistance-mediating cyp51A gene mutations from primary clinical samples, particularly as the diagnosi...
Gespeichert in:
| Hauptverfasser: | , , , , , |
|---|---|
| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
23 April 2012
|
| In: |
Antimicrobial agents and chemotherapy
Year: 2012, Jahrgang: 56, Heft: 7, Pages: 3905-3910 |
| ISSN: | 1098-6596 |
| DOI: | 10.1128/AAC.05902-11 |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1128/AAC.05902-11 Verlag, Volltext: http://aac.asm.org/content/56/7/3905 
|
| Verfasserangaben: | Birgit Spiess, Wolfgang Seifarth, Natalia Merker, Susan J. Howard, Mark Reinwald, Anne Dietz, Wolf-Karsten Hofmann, and Dieter Buchheidt |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1578242843 | ||
| 003 | DE-627 | ||
| 005 | 20220814205533.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 180803s2012 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1128/AAC.05902-11 |2 doi | |
| 035 | |a (DE-627)1578242843 | ||
| 035 | |a (DE-576)508242843 | ||
| 035 | |a (DE-599)BSZ508242843 | ||
| 035 | |a (OCoLC)1341015908 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 33 |2 sdnb | ||
| 100 | 1 | |a Spiess, Birgit |d 1972- |e VerfasserIn |0 (DE-588)1028287496 |0 (DE-627)730538699 |0 (DE-576)37578764X |4 aut | |
| 245 | 1 | 0 | |a Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients |c Birgit Spiess, Wolfgang Seifarth, Natalia Merker, Susan J. Howard, Mark Reinwald, Anne Dietz, Wolf-Karsten Hofmann, and Dieter Buchheidt |
| 264 | 1 | |c 23 April 2012 | |
| 300 | |a 6 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Published ahead of print 23 April 2012 | ||
| 500 | |a Gesehen am 03.08.2018 | ||
| 520 | |a The increasing incidence of azole resistance in Aspergillus fumigatus causing invasive aspergillosis (IA) in immunocompromised/hematological patients emphasizes the need to improve the detection of resistance-mediating cyp51A gene mutations from primary clinical samples, particularly as the diagnosis of invasive aspergillosis is rarely based on a positive culture yield in this group of patients. We generated primers from the unique sequence of the Aspergillus fumigatus cyp51A gene to establish PCR assays with consecutive DNA sequence analysis to detect and identify the A. fumigatus cyp51A tandem repeat (TR) mutation in the promoter region and the L98H and M220 alterations directly in clinical samples. After testing of the sensitivity and specificity of the assays using serially diluted A. fumigatus and human DNA, A. fumigatus cyp51A gene fragments of about 150 bp potentially carrying the mutations were amplified directly from primary clinical samples and subsequently DNA sequenced. The determined sensitivities of the PCR assays were 600 fg, 6 pg, and 4 pg of A. fumigatus DNA for the TR, L98H, and M220 mutations, respectively. There was no cross-reactivity with human genomic DNA detectable. Sequencing of the PCR amplicons for A. fumigatus wild-type DNA confirmed the cyp51A wild-type sequence, and PCR products from one azole-resistant A. fumigatus isolate showed the L98H and TR mutations. The second azole-resistant isolate revealed an M220T alteration. We consider our assay to be of high epidemiological and clinical relevance to detect azole resistance and to optimize antifungal therapy in patients with IA. | ||
| 700 | 1 | |a Seifarth, Wolfgang |d 1960- |e VerfasserIn |0 (DE-588)1042491887 |0 (DE-627)769066453 |0 (DE-576)393988899 |4 aut | |
| 700 | 1 | |a Merker, Natalia |d 1978- |e VerfasserIn |0 (DE-588)1080535055 |0 (DE-627)844777668 |0 (DE-576)453600182 |4 aut | |
| 700 | 1 | |a Reinwald, Mark |d 1975- |e VerfasserIn |0 (DE-588)131809083 |0 (DE-627)514269928 |0 (DE-576)298766515 |4 aut | |
| 700 | 1 | |a Hofmann, Wolf-Karsten |d 1967- |e VerfasserIn |0 (DE-588)114154635 |0 (DE-627)691208751 |0 (DE-576)351568336 |4 aut | |
| 700 | 1 | |a Buchheidt, Dieter |d 1955- |e VerfasserIn |0 (DE-588)1028287461 |0 (DE-627)730538621 |0 (DE-576)375787380 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Antimicrobial agents and chemotherapy |d Washington, DC : Soc., 1972 |g 56(2012), 7, Seite 3905-3910 |h Online-Ressource |w (DE-627)306316919 |w (DE-600)1496156-8 |w (DE-576)09718859X |x 1098-6596 |7 nnas |a Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients |
| 773 | 1 | 8 | |g volume:56 |g year:2012 |g number:7 |g pages:3905-3910 |g extent:6 |a Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1128/AAC.05902-11 |x Verlag |x Resolving-System |3 Volltext |
| 856 | 4 | 0 | |u http://aac.asm.org/content/56/7/3905 |x Verlag |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20180803 | ||
| 993 | |a Article | ||
| 994 | |a 2012 | ||
| 998 | |g 1028287461 |a Buchheidt, Dieter |m 1028287461:Buchheidt, Dieter |d 60000 |d 61200 |e 60000PB1028287461 |e 61200PB1028287461 |k 0/60000/ |k 1/60000/61200/ |p 8 |y j | ||
| 998 | |g 114154635 |a Hofmann, Wolf-Karsten |m 114154635:Hofmann, Wolf-Karsten |d 60000 |d 61200 |e 60000PH114154635 |e 61200PH114154635 |k 0/60000/ |k 1/60000/61200/ |p 7 | ||
| 998 | |g 131809083 |a Reinwald, Mark |m 131809083:Reinwald, Mark |d 60000 |d 61200 |e 60000PR131809083 |e 61200PR131809083 |k 0/60000/ |k 1/60000/61200/ |p 5 | ||
| 998 | |g 1080535055 |a Merker, Natalia |m 1080535055:Merker, Natalia |d 60000 |d 61200 |e 60000PM1080535055 |e 61200PM1080535055 |k 0/60000/ |k 1/60000/61200/ |p 3 | ||
| 998 | |g 1042491887 |a Seifarth, Wolfgang |m 1042491887:Seifarth, Wolfgang |d 60000 |d 61200 |e 60000PS1042491887 |e 61200PS1042491887 |k 0/60000/ |k 1/60000/61200/ |p 2 | ||
| 998 | |g 1028287496 |a Spiess, Birgit |m 1028287496:Spiess, Birgit |d 60000 |d 61200 |e 60000PS1028287496 |e 61200PS1028287496 |k 0/60000/ |k 1/60000/61200/ |p 1 |x j | ||
| 999 | |a KXP-PPN1578242843 |e 3020193079 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"note":["Published ahead of print 23 April 2012","Gesehen am 03.08.2018"],"type":{"media":"Online-Ressource","bibl":"article-journal"},"recId":"1578242843","language":["eng"],"title":[{"title":"Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients","title_sort":"Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients"}],"person":[{"roleDisplay":"VerfasserIn","display":"Spiess, Birgit","role":"aut","family":"Spiess","given":"Birgit"},{"roleDisplay":"VerfasserIn","display":"Seifarth, Wolfgang","role":"aut","family":"Seifarth","given":"Wolfgang"},{"family":"Merker","given":"Natalia","display":"Merker, Natalia","roleDisplay":"VerfasserIn","role":"aut"},{"roleDisplay":"VerfasserIn","display":"Reinwald, Mark","role":"aut","family":"Reinwald","given":"Mark"},{"role":"aut","roleDisplay":"VerfasserIn","display":"Hofmann, Wolf-Karsten","given":"Wolf-Karsten","family":"Hofmann"},{"family":"Buchheidt","given":"Dieter","display":"Buchheidt, Dieter","roleDisplay":"VerfasserIn","role":"aut"}],"physDesc":[{"extent":"6 S."}],"relHost":[{"id":{"zdb":["1496156-8"],"eki":["306316919"],"issn":["1098-6596"]},"origin":[{"publisherPlace":"Washington, DC","publisher":"Soc.","dateIssuedKey":"1972","dateIssuedDisp":"1972-"}],"name":{"displayForm":["publ. by the American Society for Microbiology"]},"physDesc":[{"extent":"Online-Ressource"}],"title":[{"title_sort":"Antimicrobial agents and chemotherapy","subtitle":"AAC","title":"Antimicrobial agents and chemotherapy"}],"part":{"pages":"3905-3910","issue":"7","year":"2012","extent":"6","volume":"56","text":"56(2012), 7, Seite 3905-3910"},"titleAlt":[{"title":"AAC"}],"pubHistory":["1.1972 -"],"language":["eng"],"corporate":[{"role":"isb","display":"American Society for Microbiology","roleDisplay":"Herausgebendes Organ"}],"recId":"306316919","type":{"media":"Online-Ressource","bibl":"periodical"},"disp":"Development of novel PCR assays to detect azole resistance-mediating mutations of the aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patientsAntimicrobial agents and chemotherapy","note":["Gesehen am 09.06.21"]}],"origin":[{"dateIssuedDisp":"23 April 2012","dateIssuedKey":"2012"}],"id":{"eki":["1578242843"],"doi":["10.1128/AAC.05902-11"]},"name":{"displayForm":["Birgit Spiess, Wolfgang Seifarth, Natalia Merker, Susan J. Howard, Mark Reinwald, Anne Dietz, Wolf-Karsten Hofmann, and Dieter Buchheidt"]}} | ||
| SRT | |a SPIESSBIRGDEVELOPMEN2320 | ||