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Reagents, methods and kits for classification of fungi and direction of anti-fungal therapy

A technology of fungi and reagents, applied in the field of reagents and kits for fungal classification and guidance of antifungal therapy, which can solve problems such as adverse side effects and expensive broad-spectrum antifungal drugs

Inactive Publication Date: 2009-06-03
ADVANDX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, broad-spectrum antifungal drugs can be expensive or associated with adverse side effects

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0140] PNA probe sequence

[0141] Fluconazole Sensitive Probe Set

[0142] Probe A-Fam Fam-OO-AGAGAGCAGCATGCA-NH 2 (Sequence ID No. 1)

[0143] Fluconazole resistance probe set

[0144] Probe B-Tam Tam-OO-GCAAGGGGCGCAAA-NH 2 (Sequence ID No. 2)

[0145] Probe F-Tam Tam-OO-GCAGCGGTGCGCAA-NH 2 (Sequence ID #6)

[0146] (Note: Conventional nomenclature used to illustrate the ends of PNA probes; O = 8-amino-3,6-dioxanoic acid; Fam = 5(6)-carboxyfluorescein, Tam = 5(6)-carboxytetramethyl kyrodamine.)

[0147] bacteria

[0148] The reference strains prepared by overnight culture presented different Candida species, including Candida albicans (C. albicans), Candida glabrata (C. .kefyr), Candida krusei (C.krusei), Candida portuguese (C.lusitaniae), Candida parapsilosis (C.parapsilosis), Candida tropicalis (C.tropicalis) and baker's yeast, Saccharomyces cerevisiae (S cerevisiae) and Staphylococcus epidermidis, common blood culture contaminants.

[0149] Smear preparat...

Embodiment 2

[0161] Example 2 was carried out in the same manner as Example 1 except that the fluconazole-sensitive probe group was added with a second probe.

[0162] PNA probe sequence

[0163] Fluconazole Sensitive Probe Set

[0164] Probe A-Fam Fam-OO-AGAGAGCAGCATGCA-NH 2 (Sequence ID No. 1)

[0165] Probe I-Fam Fam-OO-GAGAGTAACATACAA-NH 2 (Sequence ID No. 9)

[0166] Fluconazole resistance probe set

[0167] Probe B-Tam Tam-OO-GCAAGGGGCGCAAA-NH 2 (Sequence ID No. 2)

[0168] Probe F-Tam Tam-OO-GCAGCGGTGCGCAA-NH 2 (Sequence ID #6)

[0169] (Note: Conventional nomenclature used to illustrate the ends of PNA probes; O = 8-amino-3,6-dioxanoic acid; Fam = 5(6)-carboxyfluorescein, Tam = 5(6)-carboxytetramethyl kyrodamine.)

[0170] Probe I-Fam was added to the hybridization solution at 500 nM along with the other probes described in Example 1. Microscopy was performed using a fluorescence microscope equipped with a FITC / Texas Red dual-band filter. Fluconazole-sensitive fungi w...

Embodiment 3

[0179] Example 3 was performed in exactly the same manner as Example 1 except with a different probe set.

[0180] PNA probe sequence

[0181] Fluconazole Moderate Probe Set

[0182] Probe N-Tam Tam-OO-AAGAAGTAACATACA-NH 2 (Sequence ID No. 14)

[0183] (Note: Conventional nomenclature used to illustrate the ends of PNA probes; O = 8-amino-3,6-dioxanoic acid; Tam = 5(6)-carboxytetramethylrhodamine.)

[0184] Probe N-Tam was added to the hybridization solution at 250 nM. Microscopy was performed using a fluorescence microscope equipped with a FITC / Texas Red dual-band filter. Identification of fluconazole moderate fungi by red fluorescent buds. The results are reported in Table 4.

[0185] Table 4

[0186] Strain identification result incidence rate 1 Candida albicans (C.albicans) feminine 54.5％ Candida glabrata (C.glabrata) feminine 12.3％ Candida krusei (C. krusei) feminine 1.5％ Candida parapsilosis (C.parapsilosis) feminine...

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PUM

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Abstract

The present invention provides methods, kits and compositions to classify fungi. Methods are provided for classification of fungi according to established phenotypes, for example, antimicrobial susceptibility profiles. More specifically, the invention provides methods for the use of PNA probes in diagnostic applications, which will aid in the direction of appropriate therapy against fungi.

Description

[0001] Related applications [0002] This application claims priority from U.S. Provisional Application No. 60 / 719,153, filed September 20, 2005, and U.S. Provisional Application No. 60 / 789,656, filed April 5, 2006, which The entire contents of are incorporated into this case by reference. technical background [0003] Fungal infections, especially those caused by yeast, are associated with high morbidity and mortality. The increase in the prevalence of fungal infections in hospitals, especially bloodstream infections (BSIs) caused by Candida species, has resulted in an overall increase in the proportion of BSIs caused by yeast (Wisplinghoff CID 2004). [0004] The susceptibility of different Candida species to available antifungal compounds is continuously retested and the results are widely published. The susceptibility of isolates to drugs is usually described in terms of minimum inhibitory concentration units (MIC). Although the susceptibility of specific isolates varies...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor H·斯滕德M·菲安达卡
Owner ADVANDX