Mycoplasma Isolation and Identification Culture Tube (Culture Method)

Instructions for Mycoplasma Isolation and Identification Culture Tubes (Culture Method)

[Product Name] Mycoplasma Isolation and Identification Culture Tube (Culture Method)
[Packaging Specifications] Models: Ureaplasma urealyticum Isolation and Identification Culture Tube (Uu), Mycoplasma hominis Isolation and Identification Culture Tube (Mh), Mixed Mycoplasma Isolation and Identification Culture Tube (Uu-Mh); Specifications: 20 tests/box, 40 tests/box.
[Intended Use] Suitable for the culture of Ureaplasma urealyticum and Mycoplasma hominis in clinical settings.
[Principle of Assay] This product is designed based on the distinct biochemical reaction profiles and varying resistance to specific substances exhibited by different mycoplasma species, enabling direct detection of mycoplasma infections in clinical specimens. When used in conjunction with our company’s Mycoplasma Isolation, Identification, and Antimicrobial Susceptibility Test Kit, it can provide valuable guidance for clinicians in selecting appropriate therapeutic agents.
[Main Components] The product consists of a 4 mL low‑borosilicate glass vial containing a culture medium composed of mycoplasma basal solution, calf serum, yeast extract, biochemical substrates (urea and arginine), a mixture of antibiotics, and phenol red indicator, together with a butyl rubber stopper and a polyethylene plastic cap.
[Storage Conditions and Shelf Life] The kit should be stored at or below 0°C, with a shelf life of 12 months. The date of manufacture is indicated on the outer carton or label.
[Specimen Requirements] Specimens should be sent for testing immediately after collection; they may be stored at room temperature for no more than 2 hours, or at 2–8°C for no more than 5 hours.
Male: Use a sterile male swab to gently rotate and collect secretions by inserting it 2–4 cm into the urethral meatus; prostatic fluid or semen may also be collected for culture. If mycoplasma culture must be performed using urine, collect approximately 10 mL of the midstream portion of the first-morning voided sample, centrifuge at 2,000 rpm for 10 minutes, discard the supernatant, and retain about 1 mL of the pellet. Transfer 200 µL of this sediment to the culture medium; however, the positive rate for mycoplasma in urine is significantly lower than that in urethral secretions.
Women: Use a female swab to gently rotate at a depth of 1–2 cm within the cervical os, collect cervical secretions, and inoculate them onto a culture medium; amniotic fluid culture is also an option. However, urine specimens are not recommended.
[Testing Method]
1. Rewarming: Remove the required number of Mycoplasma culture tubes from the refrigerator and allow them to come to room temperature.
2. Inoculation: Remove the inner and outer caps of the culture tube. Insert the swab from the sample to be tested into the tube, rotate it several times while gently squeezing to ensure that the specimen on the swab is thoroughly immersed in the liquid medium, then discard the swab. For liquid specimens such as urinary sediment, semen, amniotic fluid, and prostatic fluid, pipette 0.2 mL and inoculate it onto the culture medium. Replace both the inner and outer caps, label the tube clearly, and mix thoroughly by shaking.
3. Incubation: Incubate at a constant temperature of 37°C ± 1°C for 24 to 72 hours, then observe the results.
[Interpretation of Test Results]
Mh results should be reported within 24–48 hours, with a maximum of 72 hours. If the culture medium changes from orange‑yellow to red and remains clear without significant turbidity, this indicates Mh growth and the result is considered Mh‑positive. If, after 72 hours of incubation, no change is observed, the result is deemed Mh‑negative. Should the medium become noticeably turbid, regardless of any color change, this signifies the presence of bacterial or yeast‑like microorganisms and should not be interpreted as positive for mycoplasma growth.
Uu results should be reported within 24 hours, with a maximum of 48 hours. If the culture medium changes from orange‑yellow to red and remains clear without significant turbidity, this indicates Uu growth and is interpreted as Uu‑positive; if no change is observed after 48 hours of incubation, the result is considered Uu‑negative; and if the medium becomes markedly turbid, regardless of any color change, this signifies the presence of bacterial or yeast‑like microorganisms and should not be reported as positive for mycoplasma growth.
Uu‑Mh results should be reported within 48 hours, with a maximum of 72 hours. If the culture medium changes from orange‑yellow to red and remains clear without significant turbidity, this indicates the growth of Uu and/or Mh, and the result is considered positive. If no change is observed in the culture medium, the result is interpreted as negative for Uu‑Mh growth. Should the medium become markedly turbid, regardless of any color change, this signifies the presence of bacterial or yeast‑like organisms and should not be reported as a positive result for mycoplasma growth.
If necessary, perform culture on a mycoplasma solid medium and examine under low magnification for the presence of “fried‑egg”–type colonies; their appearance indicates positive mycoplasma growth.

[Limitations of the Analytical Method]
1. This product should be operated and used by qualified laboratory personnel. Proper specimen collection and timely inoculation are critical to improving the positive rate and accuracy.
2. During the growth of Uu and Mh, the culture medium should remain clear and transparent, with no obvious turbidity; if turbidity occurs occasionally, it must not be reported as mycoplasma-positive.
3. Uu growth: Under normal conditions, the culture medium should change color within 24 hours; however, if the sample contains a low bacterial load, the color change may take longer than 24 hours. Mh growth: The culture medium typically changes color within 24–48 hours; if the sample has a high bacterial load, the color change may occur in less than 24 hours.
4. When it is uncertain whether contamination by extraneous microorganisms is present, a small aliquot of the culture medium may be inoculated onto blood agar plates or subjected to filtration through a 0.45-μm membrane filter followed by incubation; contamination can then be assessed based on colony growth on the blood agar plates or changes in the color of the filtrate.
5. After repeated freeze–thaw cycles, a small amount of precipitate may appear; however, once the medium is thoroughly mixed, it will not affect the experimental results.
[Product Performance Specifications]
1. Appearance and Physical Properties
The culture tubes for the isolation and identification of Ureaplasma urealyticum, Mycoplasma hominis, and mixed mycoplasmas should contain a pale yellow, clear liquid, free of sediment, clots, or flocculent material.
2. Analytical Sensitivity
The culture tubes for the isolation and identification of Ureaplasma urealyticum and Mycoplasma hominis shall each have a detection sensitivity of CCU = 10⁻⁴ when tested against the Ureaplasma urealyticum standard strain ATCC 27618 and the Mycoplasma hominis standard strain ATCC 14027, respectively. The mixed‑mycoplasma isolation and identification culture tube shall also exhibit a detection sensitivity of CCU = 10⁻⁴ when tested against the Ureaplasma urealyticum standard strain ATCC 27618 and the Mycoplasma hominis standard strain ATCC 14027.
3. Specificity Analysis
It exhibits a negative reaction against Gram-positive bacteria (Staphylococcus aureus ATCC 25923), Gram-negative bacteria (Escherichia coli ATCC 25922), and fungi (Candida albicans ATCC 10231).
[Precautions]
1. Contraindicated in women during menstruation and those with threatened abortion; also contraindicated after vaginal administration or irrigation.
2. This product is intended for in vitro use only and is for single-use only; do not use if the packaging is damaged.
3. After use, this product shall be disposed of in accordance with the regulations of the hospital or the environmental protection authorities.
4. Read the instructions carefully before use and use within the expiration date.

[Basic Information]
Registrant (or Filing Entity)/Manufacturer Name: Zhuhai Special Economic Zone Haitai Biopharmaceutical Co., Ltd.
Address: No. 18, Innovation Fourth Road, Gangwan Avenue, Tangjiawan Town, Zhuhai City, Guangdong Province Postal Code: 519085
Contact Information: Phone: 0756-3890858 Fax: 0756-3890848
Name of the after-sales service provider: Zhuhai Haitai Biopharmaceutical Co., Ltd.
Contact Information: Customer Technical Service Phone: 0756-3890835
E-mail: haitai@zhhaitai.com
Company website: www.zhhaitai.com
Production Address: No. 18, Innovation Road No. 4, Gangwan Avenue, Tangjiawan Town, Zhuhai City, Guangdong Province
Production License Number or Production Filing Certificate Number: Yue Food and Drug Administration Medical Device Production License No. 20040941
[Medical Device Registration Certificate Number/Product Technical Requirements Number]
[Date of Approval and Amendment of the Instruction Manual]