Methods and compositions for direct chemical lysis

Patent metadata

Field	Value
Publication number	US-10190152-B2
Application number	US-87460210-A
Country	US
Kind code	B2
Filing date	Sep 2, 2010
Priority date	Sep 3, 2009
Publication date	Jan 29, 2019
Grant date	Jan 29, 2019

Abstract

A direct chemical lysis composition includes an assay compatible buffer composition and an assay compatible surfactant. When combined with a specimen storage composition, such compositions prevent undesired modifications to nucleic acid and proteins lysed from cells in the biological sample. Assays of samples from such compositions do not require expensive and time-consuming steps such as centrifugation and prolonged high temperature processing. The direct chemical lysis composition of the present invention permits direct nucleic acid extraction from the cells in the biological sample without the need to decant off the transport media or otherwise exchange the transport media with assay compatible buffers. There is no need to combine the sample with proteinase K or another enzyme to extract nucleic acids from the cells. A method for lysing cells to obtain target nucleic acid for assay and a kit for combining the direct chemical lysis composition with a sample are also contemplated.

First claim

What is claimed is:

1. A composition consisting of a direct chemical lysis composition in combination with a liquid-based cytology sample or a formalin-fixed, paraffin embedded sample, the direct chemical lysis composition consisting of:
a) a buffer composition comprising a buffer component and a metal salt component, wherein the metal salt component is selected from the group consisting of sodium chloride (NaCl), potassium chloride (KCl), sodium acetate (C 2 H 3 NaO 2 ) and ammonium sulfate ((NH 4 ) 2 SO 4 ), wherein a concentration of the buffer component is in a range of 0.2 M to 2 M and a concentration of the metal salt component is in a range of 0.01 M to 1 M; and
b) a non-ionic surfactant wherein a concentration of the non-ionic surfactant is in a range of 0.01 to 2 percent (v/v).

2. The direct chemical lysis composition of claim 1 wherein the direct chemical lysis composition has a pH of about 6.6 to about 10.

3. The direct chemical lysis composition of claim 1 wherein the metal salt component is NaCl and the NaCl concentration is in the range of 0.01 M to 1 M.

4. The direct chemical lysis composition of claim 3 wherein the buffer component is selected from the group consisting of tris(hydroxymethyl)amino methane and an acid salt of tris(hydroxymethyl)amino methane.

5. The direct chemical lysis composition of claim 4 wherein the non-ionic surfactant is a polyethylene glycol based non-ionic surfactant.

6. The direct chemical lysis composition of claim 1 wherein the non-ionic surfactant is selected from the group consisting of polyoxyethylene (20) sorbitol monolaurate and polyethylene glycol octyphenyl ether.

7. The direct chemical lysis composition of claim 6 wherein the buffer component is an acid salt of tris(hydroxymethyl)amino methane and the buffer component concentration is 0.75 M, the metal salt component is NaCl and the NaCl concentration is 0.19 M and the non-ionic surfactant is polyethylene glycol octylphenyl ether and the polyethylene glycol octylphenyl ether concentration is about 0.75 percent (v/v).

Assignees

• Yang Feng
• Wang Sha Sha
• Vaughan Laurence Michael
• Porter Michael
• Rose Elaine
• Becton Dickinson Co

Inventors

• Yang Feng
• Wang Sha-Sha
• Vaughan Laurence Michael
• Porter Michael
• Rose Elaine

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• G01N2333/025

  Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus · CPC title

• C12Q1/708

  for papilloma · CPC title

• G01N33/56983

  Viruses · CPC title

• C12N1/06

  Lysis of microorganisms · CPC title