Background: Unlike the Mycoplasma IST2 kit (bioMérieux), the Mycoplasma IST3 kit has been updated to comply with the standardized antimicrobial susceptibility test (AST) method for Ureaplasma spp. (Up) and Mycoplasma hominis (Mh). We aimed to verify the use of the Mycoplasma IST3 kit for genital mycoplasma cultures. Methods: From September 2023 to January 2024, the R1 medium remaining after inoculation with IST2 was refrigerated until the next day. For IST2-positive samples, 300 μL of residual R1 medium was inoculated into the IST3. Species identification, enumeration, and AST results obtained using IST3 were compared with those obtained using IST2. Results: A total of 48 IST2-positive samples were inoculated into IST3, including 35, 1, and 12 Up-only, Mh-only, and both Up- and Mh-positive samples, respectively. Among Up-only samples, 2.8%, 91.4%, and 100.0% were susceptible to ciprofloxacin, tetracycline, and erythromycin, respectively. With IST3, 45 (93.8%) samples grew genital mycoplasmas; 42 (89.4%) of the 47 Up-positive samples and 6 (46.2%) of the 13 Mh-positive samples showed growth of the same organisms. All seven samples that failed to grow Mh were from mixed cultures, of which four Mh concentrations of < 104 /mL. Up was susceptible to levofloxacin, tetracycline, and erythromycin at the rates of 64.3 %, 88.1 %, and 95.2 %, respectively. Conclusion IST3 showed good performance in detecting genital Mycoplasma except for its tendency to not detect Mh of low concentrations in mixed cultures. IST3 is preferable to IST2 because it can accurately screen for erythromycin resistance in Up and reduce falseresistances for fluoroquinolone.

Background: Unlike the Mycoplasma IST2 kit (bioMérieux), the Mycoplasma IST3 kit has been updated to comply with the standardized antimicrobial susceptibility test (AST) method for Ureaplasma spp. (Up) and Mycoplasma hominis (Mh). We aimed to verify the use of the Mycoplasma IST3 kit for genital mycoplasma cultures. Methods: From September 2023 to January 2024, the R1 medium remaining after inoculation with IST2 was refrigerated until the next day. For IST2-positive samples, 300 μL of residual R1 medium was inoculated into the IST3. Species identification, enumeration, and AST results obtained using IST3 were compared with those obtained using IST2. Results: A total of 48 IST2-positive samples were inoculated into IST3, including 35, 1, and 12 Up-only, Mh-only, and both Up- and Mh-positive samples, respectively. Among Up-only samples, 2.8%, 91.4%, and 100.0% were susceptible to ciprofloxacin, tetracycline, and erythromycin, respectively. With IST3, 45 (93.8%) samples grew genital mycoplasmas; 42 (89.4%) of the 47 Up-positive samples and 6 (46.2%) of the 13 Mh-positive samples showed growth of the same organisms. All seven samples that failed to grow Mh were from mixed cultures, of which four Mh concentrations of < 104 /mL. Up was susceptible to levofloxacin, tetracycline, and erythromycin at the rates of 64.3 %, 88.1 %, and 95.2 %, respectively. Conclusion IST3 showed good performance in detecting genital Mycoplasma except for its tendency to not detect Mh of low concentrations in mixed cultures. IST3 is preferable to IST2 because it can accurately screen for erythromycin resistance in Up and reduce falseresistances for fluoroquinolone.

Acanthamoeba species are ubiquitous, free-living organisms found in the environment. They can cause a sight-threatening cornea disease, termed Acanthamoeba keratitis, and are often misdiagnosed, causing delayed administration of the correct treatment. Herein, we report a case of Acanthamoeba keratitis diagnosed without culture. A 12-year-old girl with a history of wearing contact lenses presented with complaints of pain, irritation, and hyperemia in the left eye. Corneal scraping-smeared slide, and liquids with contact lenses were submitted to the clinical microbiology laboratory. Cultures of Acanthamoeba spp. were not available; thus, they were stained with calcofluor white. The isolation of Acanthamoeba from the corneal scraping allowed the detection of trophozoites and cysts based on their morphological characteristics. PCR targeting the 18s rRNA gene and subsequent sequencing revealed 99% identity with the Acanthamoeba spp. Although it is challenging to find real-world evidence of Acanthamoeba in clinical microbiology without using culture methods, this case underscores the need for clinical microbiology laboratories to maintain their inspection capabilities.

Background: Unlike the Mycoplasma IST2 kit (bioMérieux), the Mycoplasma IST3 kit has been updated to comply with the standardized antimicrobial susceptibility test (AST) method for Ureaplasma spp. (Up) and Mycoplasma hominis (Mh). We aimed to verify the use of the Mycoplasma IST3 kit for genital mycoplasma cultures. Methods: From September 2023 to January 2024, the R1 medium remaining after inoculation with IST2 was refrigerated until the next day. For IST2-positive samples, 300 μL of residual R1 medium was inoculated into the IST3. Species identification, enumeration, and AST results obtained using IST3 were compared with those obtained using IST2. Results: A total of 48 IST2-positive samples were inoculated into IST3, including 35, 1, and 12 Up-only, Mh-only, and both Up- and Mh-positive samples, respectively. Among Up-only samples, 2.8%, 91.4%, and 100.0% were susceptible to ciprofloxacin, tetracycline, and erythromycin, respectively. With IST3, 45 (93.8%) samples grew genital mycoplasmas; 42 (89.4%) of the 47 Up-positive samples and 6 (46.2%) of the 13 Mh-positive samples showed growth of the same organisms. All seven samples that failed to grow Mh were from mixed cultures, of which four Mh concentrations of < 104 /mL. Up was susceptible to levofloxacin, tetracycline, and erythromycin at the rates of 64.3 %, 88.1 %, and 95.2 %, respectively. Conclusion IST3 showed good performance in detecting genital Mycoplasma except for its tendency to not detect Mh of low concentrations in mixed cultures. IST3 is preferable to IST2 because it can accurately screen for erythromycin resistance in Up and reduce falseresistances for fluoroquinolone.

Background: Unlike the Mycoplasma IST2 kit (bioMérieux), the Mycoplasma IST3 kit has been updated to comply with the standardized antimicrobial susceptibility test (AST) method for Ureaplasma spp. (Up) and Mycoplasma hominis (Mh). We aimed to verify the use of the Mycoplasma IST3 kit for genital mycoplasma cultures. Methods: From September 2023 to January 2024, the R1 medium remaining after inoculation with IST2 was refrigerated until the next day. For IST2-positive samples, 300 μL of residual R1 medium was inoculated into the IST3. Species identification, enumeration, and AST results obtained using IST3 were compared with those obtained using IST2. Results: A total of 48 IST2-positive samples were inoculated into IST3, including 35, 1, and 12 Up-only, Mh-only, and both Up- and Mh-positive samples, respectively. Among Up-only samples, 2.8%, 91.4%, and 100.0% were susceptible to ciprofloxacin, tetracycline, and erythromycin, respectively. With IST3, 45 (93.8%) samples grew genital mycoplasmas; 42 (89.4%) of the 47 Up-positive samples and 6 (46.2%) of the 13 Mh-positive samples showed growth of the same organisms. All seven samples that failed to grow Mh were from mixed cultures, of which four Mh concentrations of < 104 /mL. Up was susceptible to levofloxacin, tetracycline, and erythromycin at the rates of 64.3 %, 88.1 %, and 95.2 %, respectively. Conclusion IST3 showed good performance in detecting genital Mycoplasma except for its tendency to not detect Mh of low concentrations in mixed cultures. IST3 is preferable to IST2 because it can accurately screen for erythromycin resistance in Up and reduce falseresistances for fluoroquinolone.