PNA probes, mixtures, methods and kits pertaining to the determination of Mycoplasma and related Mollicutes

a technology of mycoplasma and mollicutes, applied in the field of detection, analysis and/or quantification of microorganisms, can solve the problems of affecting the biosynthetic ability of all species of mycoplasma, and affecting the quality and productivity of cell cultur

Inactive Publication Date: 2006-11-09
APPL BIOSYSTEMS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because of their small size, these organisms have limited biosynthetic capabilities, making them dependent on external sources for essential nutrients and cofactors.
All species of Mycoplasma are considered a risk as potential cell culture contaminants.
For example, Mycoplasma species can compete with cultured cells for essential nutrients or can produce toxins that can cause cell death, all of which can impact the quality and productivity of cell cultures.
Current culture methods for detection of Mollicutes are limited by the time required for the growth of these fastidious organisms.
As a result, detection of Mollicutes using these methods can take 28 days or longer, a timeframe that is not compatible with today's fast pace of pharmaceutical manufacturing and distribution.

Method used

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  • PNA probes, mixtures, methods and kits pertaining to the determination of Mycoplasma and related Mollicutes
  • PNA probes, mixtures, methods and kits pertaining to the determination of Mycoplasma and related Mollicutes
  • PNA probes, mixtures, methods and kits pertaining to the determination of Mycoplasma and related Mollicutes

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example 1

Detection of Mycoplasma and Related Genera

[0103] Although many polymerase chain reaction assays have been devised for the detection of Mycoplasma species and related genera of Mollicutes (Dussurget, et al., 1994; Haier et al., 1999; Jensen et al., 1999; Kidder et al., 1993; Tang et al., 2000; van Kuppeveld et al, 1992; van Kuppeveld et al., 1994), the difficulty in identifying primers that selectively amplify sequences belonging to this large and phylogenetically diverse group of organisms is reflected in the fact that none of these primer sets have been shown to be entirely specific for their intended targets (Kong et al., 2001). The combined use of DNA primers and PNA probes described here for detection of Mycoplasma and related genera is more specific than these previous assays because it contains two layers of specificity. The first layer of specificity lies in the combination of the five DNA primers described here (SEQ ID Nos. 1-5), which were selected to encompass the sequence...

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Abstract

This invention is related to PNA probes, probe sets, mixtures, methods and kits pertaining to the determination of Mycoplasma and related Mollicutes.

Description

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 60 / 678,331 filed May 6, 2005, which is incorporated herein by reference in its entirety.[0002] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described in any way.1. FIELD[0003] This invention is related to the field of detection, analysis and / or quantification of microorganisms such as for the determination of Mycoplasma and related Mollicutes.2. INTRODUCTION[0004] Members of the Class Mollicutes are among the smallest and simplest of the prokaryotes. This Class includes species from the genera Mycoplasma, Acholeplasma and Ureaplasma, and may be agents of disease and / or present as adventitious contaminants in cell culture, or in certain products and processes. Because of their small size and lack of a rigid cell wall, members of the Class Mollicutes (referred to hereafter simply as "Mollicutes") pass easily through f...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C07K14/195
CPCY10T436/143333C12Q1/6816C12Q2525/313C12Q2525/107
Inventor HYLDIG-NIELSEN, JENS J.RIGBY, SUSANBREHM-STECHER, BYRON F.LEE, DITTE S.TANNER, MICHAEL
Owner APPL BIOSYSTEMS INC
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