Method for detecting microorganisms
AssigneesNational University Corporation Tokyo Medical and Dental University, Nihon Techno Service Co., Ltd., National University Corporation Oita University, Riken, Shimadzu Corporation
InventorsNorio Shimizu, Hiroshi Takase, Manabu Mochizuki, Yasuhiro Tomaru, Satoko Nakano, Sunao Sugita, Masamitsu Shikata
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Abstract
The present invention relates to a method for simultaneously detecting a plurality of pathogens from biologically-derived samples, and a kit for carrying out the method. Specifically, the present invention relates to a method for simultaneously detecting a plurality of pathogens that cause infectious uveitis, one of eye infections from samples such as anterior chamber fluid or vitreous by polymerase chain reaction (PCR), and a kit for carrying out the method.
Claims8
A method for detecting microorganisms in a sample, comprising
- (1) a step of mixing the sample with a PCR buffer comprising a nonionic surfactant;
- Tris buffer, containing KCl and MgCl2;
- and a dNTP mix of dATP, dGTP, dCTP and dTTP to obtain a mixed solution;
- (2) a step of adding the mixed solution in step (1) to a plurality of tubes wherein each tube of the plurality of tubes comprises a solid composition for PCR reaction comprising a DNA polymerase and two types of PCR primer pairs which are selected from the group consisting of:
- (i) a Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene detection primer pair having the sequences of SEQ ID NO:1 and 2 and a TATA-binding protein (TBP) gene detection primer pair having the sequences of SEQ ID NO:3 and 4,
- (ii) a HSV-1 detection primer pair having the sequences of SEQ ID NO:
- 5 and 6 and a VZV detection primer pair having the sequences of SEQ ID NO:
- 7 and 8,
- (iii) a HSV-2 detection primer pair having the sequences of SEQ ID NO:
- 9 and 10 and a HHV-6 detection primer pair having the sequences of SEQ ID NO:
- 11 and 12,
- (iv) a EBV detection primer pair having the sequences of SEQ ID NO:
- 13 and 14 and a CMV detection primer pair having the sequences of SEQ ID NO:
- 15 and 16, and
- (v) a HTLV-1 detection primer pair having the sequences of SEQ ID NO:
- 17 and 18 and a Treponema pallidum detection primer pair having the sequences of SEQ ID NO:
- 19 and 20 and a toxoplasma detection primer pair having the sequences of SEQ ID NO:21 and 22;
- and
- (3) a step of detecting the PCR product generated in each of the plurality of tubes;
- wherein the sample is anterior chamber fluid or vitreous.
Dependencies are inferred from the claim text.