Oligonucleotide for genotyping of mycoplasma, microarray comprising the oligonucleotide, and method for detection of species using the microarray

• US 20070065828A1
• Filed: 01/14/2005
• Published: 03/22/2007
• Est. Priority Date: 01/14/2004
• Status: Abandoned Application

• Current Assignee: Genein Co., Ltd.
• Original Assignee: Genein Co., Ltd.
• Inventors: Kim, Cheol-Min, Park, Hee-Kyung, Jang, Hyun-Jung, Kim, Hyo-Myoung
• Application Number:
  US10/597,173
• Publication Number:
  US 20070065828A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/689 for bacteria

Abstract

The present invention relates to a method for detecting Mycoplasma and its related strains which are source of contamination of cell lines and biological products and human pathogenic. More particularly, the present invention relates to genus-specific and species-specific oligonucleotides for genotyping of Mycoplasma, Acholeplasm and Ureaplasma strains, microarray comprising the oligonucleotides, and method for detection of species using the microarray.

As described above, the present invention provides a rapid and accurate assay method capable of simultaneously detecting many Mycoplasma and its related strains from a single sample using a microarray comprising novel oligonucleotides for detecting Mycoplasma and its related strains which are known as a source of contamination of cell lines and biological products and human pathogenic. Further, the present invention provides an objective and credible assay method capable of tracing a contamination source for preventing expansion of infective Mycoplasma and its related strains and controlling a contamination of Mycoplasma against biological products and stem cells or cord blood cells which are useful for gene therapy and cell therapy.

Claims

1. An oligonucleotide for genus-specific genotyping of Mycoplasma and Ureaplasma strains, comprising any one sequence selected from SEQ ID Nos. 7 to 21 or its complementary sequence.

5. A microarray comprising more than one oligonucleotide selected from genus-specific and species-specific oligonucleotides for genotyping Mycoplasma, Acholeplasma and Ureaplasma strains according to any one from claims 1 to 4 as probes attached on a support.

6. The microarray according to claim 5, wherein the probes are any one selected from a group consisting of DNA, RNA, PNA, LNA and HNA.

7. The microarray according to claim 5, wherein the support is any one selected from a group consisting of slide glass, plastic, membrane, semiconductive chip, silicon and gel.

8. A method for detecting Mycoplasma strains, comprising the following steps:

a) extracting nucleic acids from a sample;

b) amplifying target DNA among the extracted nucleic acids;

c) hybridizing the amplified target DNA with probes of a microarray according to claim 5;

and

d) detecting signals generated from the hybridization reaction.

9. A kit for diagnosing Mycoplasma infection, comprising more than one oligonucleotide selected from genus-specific and species-specific oligonucleotides for genotyping Acholeplasma, Mycoplasma and Ureaplasma strains according to any one from claims 1 to 4.

2. An oligonucleotide for species-specific genotyping of Mycoplasma and Ureaplasma strains, comprising any one sequence selected from SEQ ID Nos. 28 to 127 or its complementary sequence.

3. An oligonucleotide for genus-specific genotyping of Acholeplasma strains, comprising any one sequence selected from SEQ ID Nos. 22 to 27 or its complementary sequence.

4. An oligonucleotide for species-specific genotyping of Acholeplasma strains, comprising any one sequence selected from SEQ ID Nos. 128 to 133 or its complementary sequence.