Burke B.P.,Calimmune |
Boyd M.P.,Calimmune |
Impey H.,Calimmune |
Breton L.R.,Calimmune |
And 4 more authors.
Viruses | Year: 2014
Human immunodeficiency virus type 1 (HIV-1) infection of target cells requires CD4 and a co-receptor, predominantly the chemokine receptor CCR5. CCR5-delta32 homozygosity results in a truncated protein providing natural protection against HIV infection-this without detrimental effects to the host-and transplantation of CCR5-delta32 stem cells in a patient with HIV ("Berlin patient") achieved viral eradication. As a more feasible approach gene-modification strategies are being developed to engineer cellular resistance to HIV using autologous cells. We have developed a dual therapeutic anti-HIV lentiviral vector (LVsh5/C46) that down-regulates CCR5 and inhibits HIV-1 fusion via cell surface expression of the gp41-derived peptide, C46. This construct, effective against multiple strains of both R5- and X4-tropic HIV-1, is being tested in Phase I/II trials by engineering HIV-resistant hematopoietic cells. © 2013 by the authors; licensee MDPI, Basel, Switzerland. Source
Cellex, Inc. | Date: 2015-04-13
Embodiments described herein relate to assay methods and kits for detecting resistance of any enzyme to its inhibitor due to functional alteration of the enzyme, comprising, conducting two or more reactions with two or more reagent mixes optionally containing substrates for the enzyme. The mixes are substantially similar, except that one contains no enzyme inhibitor whereas the others contain an enzyme inhibitor being tested for resistance. The ratio of the signal from the reaction with an inhibitor to that from a reaction without an inhibitor is used to indicate whether the enzyme is resistant to the enzyme inhibitor and also determine the susceptibility or resistance of the enzyme to various inhibitors and further identify enzyme variants. Embodiments further relate to assay methods comprising only two reactionsone conducted in a mix containing the inhibitor and the other without the inhibitor. Further included are devices for conducting such assays.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 2.00M | Year: 2010
DESCRIPTION (provided by applicant): The studies from our Phase I SBIR (AI082728-01) demonstrated that it is feasible to develop the proposed assay, a biochemiluminescence assay that uses a novel flu virus neuraminidase substrate, for sensitive detection of influenza virus drug resistance. The assay is now being used by a federal agency for monitoring influenza virus drug resistance during the on-going swine flu pandemic. The goal of this Phase II project is to convert the surveillance test into a drug resistance test suitable for point-of-care (POC) use (e.g., in physician's offices), i.e., a test kit that is rapid (lt 15 min for sample collection to results), simple, sensitive, specific, economic, and easy-to-use. The POC drug resistance test will also have a secondary claim for influenza diagnosis. Thus, the test is a combo test with both flu diagnosis and drug resistance detection functions. Phase II activities include GMP manufacturing, preclinical and clinical studies. The data from the Phase II studies will be submitted to the FDA for clearance of the POC test for both the influenza diagnosis and flu drug resistance detection claims. PUBLIC HEALTH RELEVANCE: Widespread resistance of seasonal influenza H1N1 to Tamiflu, the most commonly used and stockpiled drug, raises serious issue about the effectiveness of Tamiflu stockpiled for pandemic influenza, and posts a challenge for physicians who treat influenza patients. A rapid and sensitive POC flu drug resistance test would promote rational use of the drug, thereby prolonging the effectiveness of a drug, and aid the physicians in making a better prescription decision. There is no such a test available. The primary objective of the proposed Phase II studies is to build on our Phase I efforts and develop a drug resistance test that is suitable for point-of-care use.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 271.04K | Year: 2009
DESCRIPTION (provided by applicant): The primary goal of the Phase I SBIR projects is to determine the feasibility of a novel neuraminidase assay to be used as an influenza neuraminidase inhibition and drug susceptibility assay. The proposed assay is a biochemiluminescence assay that uses a novel substrate and permits real time determination of neuraminidase activity, which essentially requires only one preparation step - the addition of sample to a master detection mix that contains all necessary reagents for the assay. The proposed efforts include 1) improving substrate synthesis, 2) evaluating the feasibility as a NI and drug susceptibility assay, and 3) determining whether the assay has sufficient sensitivity so that clinical specimens can be directly used for the assay. Two neuraminidase inhibitors - zanamivir and particularly oseltamivir - are the current mainstay of pharmacological intervention during an influenza epidemic and, if happened, a pandemic. Emergence of drug resistant influenza virus variants heightens the concerns about widespread use of these drugs. It is imperative to monitor the emergence of antiviral drug resistant variants. Both cellular and biochemical based assays have been used to determine viral susceptibility to neuraminidase inhibitors. Cellular assays suffer from a number of drawbacks, including selection of mutations that compensate for neuraminidase inhibitor resistance during cell culture, long assay time, and tedious procedures, which make them less suitable for large scale surveillance programs. In contrast, the biochemical assays directly measure the inhibition of viral neuraminidase activities, thus circumventing many of the problems associated with the cellular assays. However, current biochemical assays are still complicated, take hours to complete, and have significant assay variability. More importantly, they lack sufficient sensitivity to allow drug susceptibility determination directly using clinical specimens (e.g., nasal wash), a much preferred sample type for large scale surveillance programs. Successful development of the proposed assay will allow the use of clinical specimens for drug susceptibility testing - a significant improvement over the currently used assays, which have to use virus isolates that are obtained via the lengthy and cumbersome culture methods. PUBLIC HEALTH RELEVANCE: Two neuraminidase inhibitors - zanamivir and oseltamivir - are the current mainstay of pharmacological intervention during an influenza epidemic or pandemic. Given the history of emergence of drug resistant influenza virus variants, it is imperative to monitor the emergence of antiviral drug resistant variants, which requires a robust neuraminidase inhibition (NI) assay. The proposed efforts are to determine the feasibility of a novel neuraminidase activity assay as an influenza NI assay. Specifically, we will determine if the assay is 1) rapid (5 min preparation time plus 30 min incubation); 2) simple (essentially one step operation); 3) sensitive (equivalent to or better than 25 cycles of real time PCR assay) so that clinical specimens can be directly used for NI assay; 4) reproducible (CVlt15%); and 5) suitable for a 96-well format for large-scale sample detection.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 213.34K | Year: 2016
DESCRIPTION provided by applicant A recent outbreak of carbapenem resistant Enterobacteriaceae CRE associated with contaminated duodenoscopes at the Ronald Reagan UCLA Medical Center highlights the importance of early detection of these clinically significant pathogens Coincidentally the Obama Administration released The National Action Plan to Combat Antibiotic Resistant Bacteria for which one of the steps of the action plan is to slow the emergence of resistant bacteria and prevent the spread of resistant infections Detection and identification of carbapenemase producing organisms CPO in health care centers is the first step to confining the source and preventing the potential spread of these multidrug resistant pathogens However detection of CPO in clinical laboratories is challenging as isolates may only have moderate reductions in susceptibilities to carbapenems and resistance may be mediated by other mechanisms i e chromosomal and plasmid mediated cephalosporinases or extended spectrum lactamase ESBL producers with decreased membrane permeability Molecular methods for the detection of carbapenemase genes are limited by the number of carbapenemase targets assayed in addition to other limitations The reference phenotypic method for detection of carbapenemase producing Enterobacteriaceae the Modified Hodge test suffers from a long turn around time lacks specificity and has poor sensitivity for certain types of carbapenemase enzymes i e metallo lactamase enzymes This project aims to develop a low cost easy to use essentially one manual step and rapid min assay to identify carbapenemase production in carbapenem resistant Gram negative bacteria suitable for surveillance and clinical laboratory use The proposed assay uses a luciferin derivatized lactamase substrate for detection of lactamase activity All reagents will be formulated in a master mix Reagent I such that the assay involves essentially one manual step addition of the sample to the master mix To differentiate carbapenemases from other lactamases a second master mix containing a carbapenem will be formulated Reagent II The signal of Reagent I indicates the presence or absence of lactamase while the signal ratio of Reagent II to Reagent I indicates whether the lactamase is a carbapenemase The proposed Phase I project will optimize the assay and use well characterized isolates as well as prospective clinical isolates to evaluate the assay Successful completion of this Phase I project will pave the way for a Phase II study which will involve multiple sites and substantially larger sample numbers A simple and rapid assay for detection of CPO will play a significant role in surveillance and help guide therapy and infection control measures PUBLIC HEALTH RELEVANCE A recent outbreak of carbapenem resistant Enterobacteriaceae CRE associated with contaminated duodenoscopes at the Ronald Reagan UCLA Medical Center highlights the importance of early detection of these clinically significant pathogens Detection and identification of carbapenemase producing organisms CPO in health care centers is the first step to confining the source and preventing the potential spread of these multidrug resistant pathogens The proposed project to develop a simple and rapid assay for detection of CPO if successful will play a significant role in surveillance and help guide therapy and infection control measures