COMPOSITIONS AND METHODS FOR A MYCOBACTERIUM TUBERCULOSIS DRUG SUSCEPTIBILITY TEST,申请号US201214117472-传众专利搜索

COMPOSITIONS AND METHODS FOR A MYCOBACTERIUM TUBERCULOSIS DRUG SUSCEPTIBILITY TEST

申请公布号：US2014256664(A1)

申请号：US201214117472

申请日期：2012.05.11

申请公布日期：2014.09.11

申请人：

Houpt Eric R.;Kelly Kimberly A.;Pholwat Suporn

发明人：Houpt Eric R.;Kelly Kimberly A.;Pholwat Suporn

分类号：C12Q1/68;A61K31/506;A61K31/7036;A61K31/133;A61K31/4409;A61K31/496

主分类号：C12Q1/68

地址：Keswick VA US

摘要：The present application discloses rapid Mycobacterium tuberculosis drug susceptibility utilizing real-time PCR of mycobacteriophage D29 DNA. One protocol involves culturing Tb isolates for 48 hours with and without drugs at critical concentrations, followed by incubation with 103 pfu/ml of D29 mycobacteriophage for 24 hours and then real-time PCR. Many drugs can be incubated instantly with Tb and phage. The change in phage DNA real-time PCR cycle threshold (Ct) between control Tb and Tb treated with drugs was calculated and correlated with conventional agar proportion drug susceptibility results. Specifically, 9 susceptible clinical isolates, 22 MDR, and 1 XDR Tb strains were used and Ct control−Ct drug cutoffs of between +0.3 and −6.0 yielded 422/429 (98%) accurate results for the drugs tested. The Ct values correlated with isolate minimal inhibitory concentration (MIC) for most agents. This D29 qPCR assay offers a rapid, accurate, 1-3 day phenotypic drug susceptibility test.

主权项：1. A method for determining susceptibility of Mycobacterium tuberculosis to one or more drugs, said method comprising testing a sample comprising Mycobacterium tuberculosis by: preparing at least one culture from said test sample; contacting one of said at least one cultures with a drug and optionally contacting said additional cultures of said test sample with additional drugs, wherein each additional culture receives a different drug than the other cultures; contacting said culture or additional cultures with a mycobacteriophage; determining the amount of mycobacteriophage present after contacting said culture or additional cultures with said mycobacteriophage; and comparing the amount of mycobacteriophage present in the treated culture or treated additional cultures with the amount of mycobacteriophage present in an otherwise identical untreated culture; wherein an increase in mycobacteriophage in an untreated culture relative to a culture treated with a drug is an indication that the Mycobacterium tuberculosis present in the sample is susceptible to the treatment drug, thereby determining susceptibility of Mycobacterium tuberculosis to one or more drugs.