Automated high-throughput flow-through real-time diagnostic system

Abstract

An automated real-time flow-through system capable of processing multiple samples in an asynchronous, simultaneous, and parallel fashion for nucleic acid extraction and purification, followed by assay assembly, genetic amplification, multiplex detection, analysis, and decontamination. The system is able to hold and access an unlimited number of fluorescent reagents that may be used to screen samples for the presence of specific sequences. The apparatus works by associating extracted and purified sample with a series of reagent plugs that have been formed in a flow channel and delivered to a flow-through real-time amplification detector that has a multiplicity of optical windows, to which the sample-reagent plugs are placed in an operative position. The diagnostic apparatus includes sample multi-position valves, a master sample multi-position valve, a master reagent multi-position valve, reagent multi-position valves, and an optical amplification/detection system.

1 . A method of detecting sequences in a sample using a detector with a multiplicity of optical windows, comprising the steps of:
 providing a flow channel,   forming a series of reagent plugs in said flow channel,   associating sample preparation fluids with the sample,   associating the sample with the reagent plugs forming sample-reagent plugs,   positioning said sample-reagent plugs in operative position to the multiplicity of optical windows in the detector for detecting different genetic sequences in a sample,   amplifying said sample-reagent plugs, and   detecting the sequences in the sample in said sample-reagent plugs.   
     
     
         2 . The method of detecting sequences in a sample of  claim 1  including the step of detecting sequences in additional samples by
 forming an additional series of additional reagent plugs in additional flow channels, 
 associating additional sample preparation fluids with the additional samples, 
 associating the additional samples with the additional reagent plugs forming additional sample-reagent plugs, 
 positioning said additional sample-reagent plugs in operative position to the multiplicity of optical windows in the detector for detecting different genetic sequences in the additional samples, 
 amplifying said additional sample-reagent plugs, and 
 detecting the sequences in the additional samples in said additional sample-reagent plugs. 
 
     
     
         3 . The method of detecting sequences in a sample of  claim 2  wherein said steps of
 positioning said sample-reagent plugs in operative position to the multiplicity of optical windows in the detector for detecting different genetic sequences in a sample, 
 detecting the sequences in the sample in said sample-reagent plugs, 
 positioning said additional sample-reagent plugs in operative position to the multiplicity of optical windows in the detector for detecting different genetic sequences in the additional samples, and 
 detecting the sequences in the additional samples in said additional sample-reagent plugs are performed asynchronously, simultaneously, and in parallel. 
 
     
     
         4 . The method of detecting sequences in a sample of  claim 1  including the step of associating automated nucleic acid extraction prior to assembling sample with the reagent plugs forming sample-reagent plugs. 
     
     
         5 . The method of detecting sequences in a sample of  claim 1  including the step of using decontamination fluids for decontaminating said flow channel and said detector.