Claims for Patent: 7,371,531

Summary for Patent: 7,371,531

Title:	Use of an extraction control in a method of extracting nucleic acids
Abstract:	The present invention relates to a method of ensuring the effectiveness of the extraction workup from a biological sample of nucleic acid. The inventive method is able to distinguish between possible defects in the extraction of nucleic acid from a sample and possible defects in a subsequent amplification step. The present invention also relates to a packaged array for extracting nucleic acid from a biological sample.
Inventor(s):	Hellyer; Tobin (Westminster, MD), Fort; Thomas (Hanover, PA), McMillian; Ray A. (Timonium, MD)
Assignee:	Becton, Dickinson and Company (Franklin Lakes, NJ)
Application Number:	11/114,952
Patent Claims:	1. A method of detecting, verifying, and/or calibrating the extraction of nucleic acid from a biological sample, the method comprising: a) obtaining a biological sample; b) combining one or more reference nucleic acids with the biological sample; c) extracting nucleic acid from the biological sample concurrently with and/or after step b to yield an extract; and d) detecting the presence of and/or quantifying the amount of reference nucleic acid in the extract; e) estimating the amount of non-reference nucleic acid present in the composition before the extraction step by i) quantifying the amount of total nucleic acid and the amount of reference nucleic acid in the extract of step c; ii) comparing the amount of reference nucleic acid in the extract of step c to the amount of reference nucleic acid combined with the biological sample in step a to calculate a ratio of extraction efficiency; and iii) subtracting the amount of reference nucleic acid in said extract from the amount of total nucleic in said extract to yield the amount of non-reference nucleic acid in said extract; and iv) dividing the amount of non-reference nucleic acid in the extract by the ratio of extraction efficiency, wherein the reference nucleic acid is capable of being detected and/or quantified without an amplification step, and further wherein the reference nucleic acid is capable of being detected without interference from material in the biological sample. 2. The method of claim 1, wherein the biological sample is body fluid from a human selected from the group consisting of urine, saliva, whole blood, plasma, lymph fluid, semen, vaginal fluid, sweat, tears, and mixtures thereof. 3. The method of claim 1, wherein quantifying the amount of reference nucleic acid in the extract of step c comprises: amplifying the reference nucleic acid, quantifying the amount of amplified nucleic acid, and calculating the amount of reference nucleic acid in the extract by multiplying the amplified amount of reference nucleic acid by a ratio of amplification efficiency. 4. The method of claim 1, wherein the reference nucleic acid comprises RNA or DNA labeled with a detectable moiety selected from the group consisting of a chromophore, a fluorophor, a radiolabel, or is enzyme, or is detectable by hybridization. 5. The method of claim 4, wherein the reference nucleic acid has a 3' end modified to essentially prohibit enzymatic chain extension. 6. The method of claim 4, wherein the reference nucleic acid contains a nucleic acid sequence selected from a known intron sequence and/or is capable of forming an internal hairpin structure. 7. The method of claim 1, wherein step c comprises either c1) reversibly binding nucleic acid to a solid support, washing the solid support to remove molecules that are bound less tightly than nucleic acid, and releasing nucleic acid from the solid support; or c2) irreversibly binding nucleic acid to a solid support, and washing the solid support to remove molecules that are bound less tightly than nucleic acid. 8. The method of claim 1, wherein the biological sample contains nucleic acid obtained from an organism selected from the group consisting of Chlamydia trachomatis and Neisseria gonorrhoeae. 9. The method of claim 1, wherein the reference nucleic acid comprises a set of two or more different nucleic acids that can be detected and/or quantified independently of each other. 10. A method of detecting, verifying, and/or calibrating the extraction of nucleic acid from a biological sample, the method comprising: a) obtaining a biological sample; b) combining one or more reference nucleic acids with the biological sample; c) extracting nucleic acid from the biological sample concurrently with and/or after step b to yield an extract; and d) detecting the presence of and/or quantifying the amount of reference nucleic acid in the extract; wherein the reference nucleic acid is capable of being detected and/or quantified without an amplification step, and further wherein the reference nucleic acid is capable of being detected without interference from material in the biological sample; and further wherein the reference nucleic acid is SEQ. ID NO. 1 and/or SEQ. ID NO. 2. 11. A method of analyzing a biological sample comprising: a) obtaining a biological sample; b) combining one or more reference nucleic acids with the biological sample, wherein the reference nucleic acid is capable of being detected and/or quantified without an amplification step, and further wherein the reference nucleic acid is capable of being detected without interference from material in the biological sample; c) extracting nucleic acid from the biological sample concurrently with and/or after step b to yield an extract; and d) detecting the presence of and/or quantifying the amount of reference nucleic acid in the extract; e) calculating an extraction efficiency ratio by dividing the amount of extracted reference nucleic acid by the amount of original reference nucleic acid, wherein the extraction efficiency ratio is greater than zero; and f) assaying the extract for a target nucleic acid. 12. The method of claim 11, wherein the target nucleic acid is indicative of a disease state or condition. 13. The method of claim 11, wherein the target nucleic acid is from an organism selected from the group consisting of Chlamydia trachomatis and Neisseria gonorrhoeae, and wherein the assay in step f is based on the presence of a primary structure selected from the group consisting of a structural gene, open reading frame, intron, exon, mRNA, cDNA or oligonucleotide sequence, and further wherein the reference nucleic acid is a truncated version of the target nucleic acid and contains about 50% or more of said primary structure. 14. The method of claim 11, wherein the reference nucleic acid is SEQ. ID NO. 1 and/or SEQ. ID NO. 2. 15. A packaged array for extracting nucleic acid from multiple biological samples, wherein the packaged array comprises: a) a set of first vessels containing reference nucleic acid, wherein the reference nucleic acid is capable of being detected and/or quantified without interference from target nucleic acid, and further wherein the detection and/or quantification is through a labeled moiety attached to the reference nucleic acid or through hybridization without amplification of the reference nucleic acid to a labeled species; and b) a means for extracting nucleic acid; and c) optionally a set of second vessels for independent placement of multiple biological samples that allows for transfer of material between the first vessels and the second vessels wherein the reference nucleic acid is SEQ. ID NO. 1 and/or SEQ. ID NO. 2. 16. The packaged array of claim 15, wherein the reference nucleic acid is in a dry form suitable for combination with a biological sample and subsequent extraction. 17. The packaged array of claim 15, further comprising aliquots of a solid support having an affinity for nucleic acid located in the first vessels with the reference nucleic acid or in the optional second set of vessels for placement of multiple biological samples. 18. The packaged array of claim 15, wherein the multiple biological samples are to be tested for the presence of one or more nucleic acids obtained from an organism selected from the group consisting of Chlamydia trachomatis and Neisseria gonorrhoeae.

Details for Patent 7,371,531

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2024-04-23
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2024-04-23
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2024-04-23