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Composition and kit for simultaneously detecting mycoplasma urealytium, mycoplasma hominis and mycoplasma genitalium

A technology for Ureaplasma urealyticum and Mycoplasma hominis, applied in the field of compositions and kits for detecting Mycoplasma, can solve the problem of reducing the cost of detection, inability to accurately detect Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium simultaneously, and reducing the operation time of operators and other problems, to reduce the cost of testing, improve the early detection rate, and reduce the economic burden of patients.

Active Publication Date: 2010-09-08
北京鑫诺美迪基因检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the existing composition and kit for detecting Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium. , providing a composition for simultaneous detection of Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium, using the composition can realize accurate detection of Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium simultaneously in the same reaction system, and there is no Defects that interfere with each other, reducing the operator's operating time and reducing inspection costs

Method used

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  • Composition and kit for simultaneously detecting mycoplasma urealytium, mycoplasma hominis and mycoplasma genitalium
  • Composition and kit for simultaneously detecting mycoplasma urealytium, mycoplasma hominis and mycoplasma genitalium
  • Composition and kit for simultaneously detecting mycoplasma urealytium, mycoplasma hominis and mycoplasma genitalium

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0239] A 35-year-old male patient developed urethral scab, dysuria, prostatitis and epididymitis symptoms, without other diseases. With the informed consent of the patients, the following samples were obtained:

[0240] (1) Urine sample: collect 10 ml of total urine once, centrifuge at 2000 rpm for 20 minutes, discard the supernatant, and collect the sediment. Transfer the sediment to a 1.5ml sterile tube containing 500μl normal saline for later use.

[0241] (2) Prostatic fluid sample: 10 μl of prostatic fluid was taken and stored in a 1.5 ml sterile tube filled with 500 μl of normal saline for later use.

[0242] According to "Li Jinming, real-time fluorescent PCR technology, People's Military Medical Publishing House, 2007", the method recorded in Chapter 4, Section 4, Page 73-74 respectively extracts the total DNA of the above two samples to obtain two kinds of total DNA solutions. 20 μl each.

[0243] According to the method described in "Yoshida, T., S. Maeda, T. Degu...

preparation example 2

[0245] A 27-year-old female patient developed symptoms of vaginitis and cervicitis without other complications. With the informed consent of the patients, the following cervical secretion samples were obtained:

[0246] Deepen the sterile swab about 1 cm into the cervical canal, rotate it 10 times, and stay for about 30 seconds. After taking it out, place the swab in a 1.5ml sterile tube containing 500 μl of normal saline, squeeze it fully, and discard the swab after shaking , keep 500 μl of the washing solution containing the specimen.

[0247] According to "Li Jinming, real-time fluorescent PCR technology, People's Military Medical Publishing House, 2007", the method described in chapter 4, section 4, pages 73-74, was used to extract the total DNA of the above samples to obtain 20 μl of the total DNA solution.

[0248] According to the method described in "Yoshida, T., S. Maeda, T. Deguchi, and H. Ishiko. 2002. Phylogeny-based rapid identification of mycoplasmas and ureapla...

preparation example 3

[0250] The ribosomal 16S rRNA gene of Ureaplasma urealyticum (Genbank Accession: L08642.1), the ribosomal 16S rRNA gene of Mycoplasma hominis (Genbank Accession: M96660.1) and the ribosomal 16S rRNA gene of Mycoplasma genitalium were synthesized by the solid-phase phosphite triester method (Genbank Accession: X77334.1). The obtained three DNA sequences were directly connected to the pEASY-Blunt Cloning Vector plasmid (blunt-end cloning vector) by T4 phage DNA ligase, transformed into Escherichia coli, cultivated in large quantities, and extracted the plasmid to make 10 6 copies / mL of the three simulated samples to be tested are set aside.

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Abstract

The invention discloses a composition and a kit for simultaneously detecting mycoplasma urealytium, mycoplasma hominis and mycoplasma genitalium, which comprise sequences shown by relevant SEQ ID No:1-6 or at least 15 nucleotide continuous fragments thereof, or the specific primers of sequences which have at most 5 nucleotide differences with the sequences shown by relevant SEQ ID No:1-6 or continuous fragments and sequences shown by SEQ ID No:7-9 or complementary sequences thereof or at least 15 nucleotide continuous fragments thereof or the specific probes of sequences which have at most 5 nucleotide differences with the. sequences shown by SEQ ID No:7-9 or the complementary sequences thereof or the 15 nucleotide continuous fragments The composition and the kit for simultaneously detecting the mycoplasma urealytium, the mycoplasma hominis and the mycoplasma genitalium can simultaneously detect three different mycoplasmas in one reaction system, and thereby, the early detection rate of the infectious diseases of urogenital systems is increased, the operation time of operation staffs is reduced, the detection cost is reduced and the financial burdens are relieved for patients.

Description

technical field [0001] The present invention relates to a composition and a kit for detecting mycoplasma, more particularly, the present invention relates to a composition and a kit for simultaneously detecting ureaplasma, mycoplasma hominis and mycoplasma genitalium. Background technique [0002] Ureaplasma urealyticum (UU), Mycoplasmahominis (MH) and Mycoplasma genitalium (MG) are common parasites that cause genitourinary infection in the population. Mycoplasma infection in the human urogenital tract can not only cause urogenital lesions, such as prostatitis, epididymitis, vaginitis, cervicitis, etc., but also invade the lymph nodes of the urogenital organs, and invade all important tissues and organs of the body through hematogenous spread. Organs, causing neonatal meningitis, congenital pneumonia, infertility, miscarriage, premature birth, stillbirth, low birth weight and other phenomena. Therefore, a series of lesions caused by mycoplasma infection in the human urogeni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/35C12R1/01
Inventor 叶锋赵洪斌
Owner 北京鑫诺美迪基因检测技术有限公司
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