Human Genetic Signatures Pty. Ltd.

Commonwealth Scientific and Industrial Research Organisation

Douglas Spencer Millar

George L Gabor Miklos

John R Melki

Peter Laurence Molloy

Susan Joy Clark

Cassandra Jean Vockler

Neralie Ann Coulston

Geoffrey Walter Grigg

John Robert Melki

George L Miklos

George Gabor Miklos

George Gabor L Miklos

A molecular detection assay including treating a biological sample directly with a bisulphite agent under conditions that allow cell disruption and nucleic acid treatment; removing the bisulphite agent from the treated sample; and detecting a target nucleic acid in the treated sample.

Molecular detection assay using direct treatment with a bisulphite reagent

The invention relates to the use of enzymes for copying or amplifying bisulphite modified or treated nucleic acids, wherein the enzymes are more effective in copying or amplifying the nucleic acid compared with native Taq polymerase under substantially the same conditions.

Enzymes for amplification and copying bisulphite modified nucleic acids

A method for detecting a microorganism comprising reducing the complexity of a microbial genome or microbial nucleic acid by generating a simplified form of the microbial genome or microbial nucleic acid in which substantially all of the positions naturally occupied by cytosines are occupied by uracil or thymine; and assaying for a microbial specific nucleic acid in the simplified form of the microbial genome or microbial nucleic acid, wherein presence of the microbial specific nucleic acid is indicative of the microorganism.

Methods for simplifying microbial nucleic acids by chemical modification of cytosines

The invention is directed to a method for isothermal DNA amplification comprising providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing a non-regular base, a second primer at least partially complementary to a region of DNA and containing a non- regular base, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises a non-regular base in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near the non-regular base; and amplifying the DNA substantially without thermal cycling.

Isothermal strand displacement amplification using primers containing a non-regular base

The invention relates to an assay for determining a health state of a subject using a combination of detecting the presence of a virus and detecting the presence of a genomic target or marker indicative of a health state.

Assay for screening for potential cervical cancer

Methods for treating nucleic acid including: (a) providing an alkali environment to a nucleic acid sample; (b) reacting the nucleic acid sample with a bisulphite reagent and incubating the reaction so as to form a treated nucleic acid sample where methylated nucleotides in the nucleic acid sample remain unchanged while unmethylated nucleotides are converted to another form; (c) removing unwanted reagents or diluents from the treated nucleic acid sample; and (d) carrying out de-sulphonation of the precipitated treated nucleic acid at a temperature from 70&#xb0; C. to 95&#xb0; C. by adjusting the precipitated treated nucleic acid to a pH of between 10 and less than 12.5 to remove sulphonate groups present on the treated nucleic acid and obtain a nucleic acid sample substantially free of sulphonate groups.

Bisulphite reagent treatment of nucleic acid

A diagnostic or prognostic assay is disclosed for a disease of condition characterized by abnormal methylation of cytosine at side or sites within the glutathione-S-transferase (GST) Pi gene and/or its regulatory flanking sequences (e.g., prostate cancer and liver cancer). The assay comprises: (i) isolating DNA from said subject, and (ii) determining (e.g., by selective PCR amplification) the presence of abnormal methylation of cytosine at a site or sites within the GST-Pi gene and/or its regulatory flanking sequences.

Assay for methylation in the GST-Pi gene

A method for determining the methylation status of a potential methylation site in genomic nucleic acid comprising treating genomic nucleic acid with an agent which modifies cytosine bases but does not modify 5methyl-cytosine bases under conditions to form a. modified nucleic acid template containing a potential methylation site; providing a first clamp containing a first capture sequence complementary to a region flanking one side of the potential methylation site in the modified nucleic acid template, providing a second clamp containing a second capture sequence complementary to a region flanking the other side of the potential methylation site in the modified nucleic acid template; allowing the first clamp and the second clamp to hybridise to the modified nucleic acid template; ligating the hybridised first and second clamps to form a probe spanning the potential methylation site in the modified nucleic acid template; digesting the modified nucleic acid template to obtain the probe; and detecting the probe and determining the methylation status of the potential methylation site in the modified genomic nucleic acid.

Nucleic acid detection assay

A method for simplification of a microbial genome or microbial nucleic acid comprising treating microbial genome or nucleic acid with an agent that modifies cytosine to form derivative microbial nucleic acid and amplifying the derivative microbial nucleic acid to produce a simplified form of the microbial genome or nucleic acid.

An amplification blocker comprising an intercalating nucleic acid (INA) containing two or more internal intercalating pseudonucleotides (IPNs) capable of blocking or reducing nucleic acid amplification. Use of the amplification blocker to block or reduce nucleic acid amplification.

Amplification blocker comprising intercalating nucleic acids (INA) containing intercalating pseudonucleotides (IPN)

A method for whole genome amplification comprising (a) treating genomic DNA with a modifying agent which modifies cytosine bases but does not modify 5'-methyl-cytosine bases under conditions to form single stranded modified DNA; (b) providing a population of random X-mers of exonuclease-resistant primers capable of binding to at least one strand of the modified DNA, wherein X is an integer 3 or greater; (c) providing polymerase capable of amplifying double stranded DNA, together with nucleotides and optionally any suitable buffers or diluents to the modified DNA; and (d) allowing the polymerase to amplify the modified DNA.

Methods for genome amplification

The invention provides methods for treating methylated nucleic acids. In one embodiment, the method can include the steps of (a) providing an alkali environment to a nucleic acid sample; (b) reacting the nucleic acid sample with a bisulphite reagent and incubating the reaction so as to form a treated nucleic acid sample where methylated nucleotides in the nucleic acid sample remain unchanged while unmethylated nucleotides are converted to another form; (c) diluting the treated nucleic acid sample so as to reduce salt concentration to a level which will not substantially interfere with a nucleic acid precipitating step; (d) precipitating the diluted treated nucleic acid to substantially remove any unwanted reagents or diluents from treated nucleic acid; and (e) carrying out de-sulphonation of the precipitated treated nucleic acid so as to remove sulphonate groups present on the treated nucleic acid so as to obtain a nucleic acid sample substantially free of sulphonate groups.

Growing community of inventors

Revesby, Australia

Douglas Spencer Millar