[특허]Methods for determining microbial contamination of allograft products

[미국특허] Methods for determining microbial contamination of allograft products 원문보기

IPC분류정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	C12Q-001/04 A61L-002/025 A61L-002/00
출원번호	US-0445057 (2012-04-12)
등록번호	US-9186426 (2015-11-17)
발명자 / 주소	Ronholdt, Chad J.
출원인 / 주소	Allosource
대리인 / 주소	Sheridan, James A.
인용정보	피인용 횟수 : 0 인용 특허 : 38

초록 ▼

A method for determining microbial contamination of allograft products comprises providing an extraction vessel having an outer container and a removable inner basket, place the allograft product into the inner basket, providing an abstraction fluid in the outer container of the extraction vessel, positioning the inner basket in the outer container, so that the allograft product and the inner basket is submerged in the extraction fluid and agitating the extraction fluid for a predetermined amount of time and analyzing the extraction fluid from microbial contamination.

대표청구항 ▼

1. A method of determining microbial contamination of an allograft product having a tissue matrix, comprising: providing an extraction vessel, wherein the extraction vessel comprises an outer container and a removable foraminous inner container;placing the allograft product into the inner container;providing an extraction fluid, in a given volume sufficient to fully immerse the allograft product, in the outer container of the extraction vessel;positioning the inner container in the outer container, wherein the allograft product in the inner container is submerged in the extraction fluid;agitating the extraction fluid for a predetermined amount of time by sonicating the extraction fluid at a frequency and duration sufficient to dislodge any microbial contaminants from the tissue matrix of the allograft product while maintaining the viability of any microbial contaminants dislodged from the allograft product and while maintaining the integrity of the tissue matrix of the allograft product without diminishing biomechanical properties of the tissue matrix of the allograft product;removing the inner container containing the allograft product from the outer container;filtering an entire amount of the given volume of the extraction fluid with a filter membrane to produce an extract having a higher concentration of the any microbial contaminants dislodged from the allograft product than the extraction fluid; andanalyzing the extract filtered from the extraction fluid for microbial contamination. 2. The method of claim 1, wherein analyzing the extraction fluid comprises culturing the extraction fluid. 3. A method of determining contamination of allograft products having a tissue matrix, comprising: (a) agitating an extraction fluid in which the allograft products are immersed by sonicating the extraction fluid at a frequency and duration sufficient to dislodge any microbial contaminants from the tissue matrix of the allograft product while maintaining the viability of any microbial contaminants dislodged from the allograft product and while maintaining the integrity of the tissue matrix of the allograft product without diminishing biomechanical properties of the tissue matrix of the allograft product;(b) removing the allograft products from said extraction fluid;(c) filtering an entire amount of the extraction fluid with a filter membrane to produce an extract having a higher concentration of the any microbial contaminants dislodged from the allograft product than the extraction fluid; and(d) analyzing the extract filtered from the extraction fluid for contamination. 4. The method of claim 3, wherein: during said agitating step, said extraction fluid is contained within an extraction vessel comprising an outer container and a foraminous inner container positioned within said outer container;during said agitating step, said allograft products are located within said inner container; andsaid step of removing the allograft products from said extraction fluid comprises removing said inner container from said outer container. 5. The method of claim 4, wherein said agitating step comprises manually agitating said extraction fluid. 6. The method of claim 4, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at an intensity of at least about 50 Watts per gallon. 7. The method of claim 6, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at an intensity ranging from about 50 to about 200 Watts per gallon. 8. The method of claim 6, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at an intensity ranging from about 100 to about 550 Watts per gallon. 9. The method of claim 6, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at an intensity ranging from about 200 to about 400 Watts per gallon. 10. The method of claim 4, wherein said agitating step comprises sonicating said extraction fluid at a frequency and duration sufficient to dislodge contaminants from the allograft products while maintaining the viability of any microbial contaminants dislodged from the allograft products. 11. The method of claim 10, wherein said inner container comprises a cylindrical vessel having a plurality of apertures distributed about its surface. 12. The method of claim 10, wherein said inner container comprises a mesh bag. 13. The method of claim 10, wherein said sonicating step comprises: (a) providing a sonication bath having a first fluid therein;(b) providing a sonication chamber having a second fluid therein, said sonication chamber being sized and configured to receive said outer container therein, and wherein said sonication chamber is positioned at least partially within said first fluid of said sonication bath;(c) positioning said outer container within said sonication chamber, such that the level of said second fluid within the sonication chamber is greater than the level of said extraction fluid within the outer container; and(d) applying ultrasonic energy to said first fluid in said sonication bath such that the extraction fluid within said outer container is indirectly sonicated. 14. The method of claim 4, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at a frequency of at least about 39 kHz for at least about 1 minute. 15. The method of claim 14, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at a frequency ranging from about 39 kHz to about 1 MHz. 16. The method of claim 14, wherein said agitating step comprises sonicating said extraction fluid by applying ultrasonic energy at a frequency ranging from about 39 kHz to about 172 kHz. 17. The method of claim 3, wherein said step of analyzing the extraction fluid for contamination comprises culturing said extraction fluid under conditions suitable for encouraging microbial growth. 18. The method of claim 3, wherein said step of analyzing the extraction fluid for contamination comprises culturing the extract filtered from the extraction fluid.

이 특허에 인용된 특허 (38) 인용/피인용 타임라인 분석

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[미국특허] Methods for determining microbial contamination of allograft products 원문보기