Super fast tuberculosis diagnosis and sensitivity testing method

An apparatus for mycobacterum antibiotic sensitivity testing comprising:

• (a) a means for using a mycobacterum vector to insert a marker that causes the luminescencing of individual Mycobacterium cells;
• (b) a means for locating and viewing under a luminescence detecting microscope said Mycobacterium cells;
• (c) a means for detecting luminescence in each intact Mycobacterium cell;
• (d) a means for counting the number of luminescing said Mycobacterium cells to assess whether or not there is a positive diagnosis of Mycobacterium induced diseases;
• (e) a means for recording coordinates of each of said luminescing Mycobacterium cells;
• (f) a means for segregating said luminescing Mycobacterium cells in groups;
• (g) a means for exposing said groups to an antibiotic, wherein a different antibiotic is used for each group;
• (h) a means for segregating said individual Mycobacterium cells of a said group for viewing under said microscope to determine if each of said Mycobacterium cells is luminseing;
• (i) a means to assess for each of said group, sensitivity or resistance of the said Mycobacterium in the group to the said antibiotic, based on a count of the number of said Mycobacterium cells of the group that remain continue to luminesce after a predetermined time depending on the antibiotic being tested.

A method for determining Mycobacterium antibiotic sensitivity testing comprising:

• (a) inserting a marker that causes the luminescencing of individual Mycobacterium cells using a mycobacterum vector;
• (b) locating and viewing under a luminescence detecting microscope said marked Mycobacterium cells;
• (c) detecting luminescence in each intact Mycobacterium cell;
• (d) counting the number of luminescing said Mycobacterium cells to assess whether or not there is a positive diagnosis of Mycobacterium induced diseases;
• (e) recording coordinates of each of said luminescing Mycobacterium cells;
• (f) segregating said luminescing Mycobacterium cells in groups;
• (g) exposing said groups to an antibiotic, wherein a different antibiotic is used for each group;
• (h) segregating said individual Mycobacterium cells of a said group for viewing under said microscope to determine if each Mycobacterium cells is luminseing;
• (i) assessing for each group, sensitivity or resistance of the said Mycobacterium cells in said group to the said antibiotic, based on a count of the number of said Mycobacterium cells of said group that continue to luminesce after a predetermined time depending on the antibiotic being tested.

An automated pattern recognition apparatus for recognizing mycobacterium in the field of view comprising:

• (a) an imaging means;
• (b) a scanning means using a TV camera or a line scan camera whereby the image intensity profiles of the scanning lines are recorded in a computers memory;
• (c) a processing means;
• wherein the darker portions of said profiles are distinguished as covers;
• wherein the coordinates of the beginning and ending of the said covers is ascertained and recorded in a computer's memory;
• wherein the relationship between the said beginning and ending of said covers from successive said scanning lines is analyzed;
• and
• wherein said analysis consists of a means for distinguishing between said relationships that represent mycobacterium images and images of non- mycobacterium objects in the image.

An apparatus for improving the efficiency of the electroporesis process on a specimen of suspended microorganisms comprising:

• (a) a chamber, the interior of which is suitable for holding a specimen and the exterior of which is suitable for being placing on a microscope stage;
• (b) a plurality of electrodes energisable by at least one power supply;
• wherein said electrodes when energized produce an electrostatic field in the fluid filled part of said chamber that rotates microorganisms so their long axis are perpendicular to the surface of the electrodes;
• wherein said electrodes when energized produce electroporesis electrical pulses for inserting the plasmid into said microorganisms;
• (c) an electric circuit suitable for producing the said electrostatic field and the said electroporesis electrical pulses.

An apparatus for effectuating the fast Mycobacterium diagnosis and sensitivity testing comprising:

• (a) a computer controlled automated microscope for viewing and analyzing microorganisms in the field of view comprising:
• (b) a high power microscope objective lens for viewing a field of view consisting of one of the following:
• (1) a 100.times.objective;
• (2) a phase 100.times.objective;
• (c) a light source for illuminating the field of view that can be turned on or off;
• (d) a computer controlled motorized microscope stage capable of holding a chamber on said stage;
• (e) a means of moving a microorganism by automated means under the said objective lens, then automatically determining the coordinates of the stage corresponding to the microorganism and storing said coordinates in a computer's memory;
• (f) a means for automatically relocating said respective microorganisms under the said objective lens by retrieving the coordinates from the computer's memory and then repositioning the said stage to the said corresponding coordinates.

An apparatus for enabling both immunofluorescence followed by luminescence to be used for identifying a microorganism and then for antibiotic sensitivity testing of microorganisms comprising:

• (a) a chamber of a size suitable for being placing on a microscope stage shaped with flat top and bottom sides;
• wherein one of said sides is transparent for viewing a specimen under a microscope and at least one side is transparent/translucent to enable light to illuminate the said specimen;
• wherein the sides of said chamber form an interior which is subdivided into a plurality of group modules;
• wherein a portion of the said sample can be held in each module, so that simultaneously each module can be used to test the sensitivity of the microorganisms to an antibiotic different from that tested in the other modules.