Comparison of gradient diffusion and molecular methods using Allplex™ NG&DR assay (Seegene®) for macrolide and fluoroquinolone screening resistance in Neisseria gonorrhoeae.

Antimicrobial resistance in Neisseria gonorrhoeae (NG) is increasing worldwide. Second-line treatments with macrolides or fluoroquinolones are an option for NG infections in some cases following the STI guideline recommendations. In our study, we compared the gradient diffusion test using EUCAST 2024 breakpoints with a new molecular method using the Allplex™ NG&DR assay (Seegene®) including A2059G/C2611 mutations (23S rRNA) associated with high/moderate-level macrolide resistance and S91F mutation (gyrA) relationship with fluoroquinolone resistance in NG isolates (n = 100). We calculated the sensitivity, specificity, and correlation of the molecular test for fluoroquinolone using the gradient diffusion as the reference method. In twenty-three strains was not detected any mutation associated with macrolides or fluoroquinolone resistance. No A2059G/C2611T mutations were detected, and the S91F mutations were detected in 77 out of the 100 isolates screened. Twenty-three NG isolates were reported to be resistant to azithromycin (ECOFF: >1 mg/L), and 78 NG isolates were resistant to ciprofloxacin (MIC: >0.06 mg/L). The molecular method showed a sensitivity of 96.1% and, a specificity of 90.9% for fluoroquinolone susceptibility, but the statistical analysis between the molecular test and gradient diffusion test was not statistically significant for fluoroquinolone resistance (p = 1). Statistical analysis was not performed for macrolides because of the absence of positive RT-PCR results. According to our data, Allplex™ assay cannot replace the gradient diffusion test for macrolide resistance. However, the assay could be used to test fluoroquinolone resistance in NG isolates as a replacement for phenotypic methods.

Increase in gonococcal arthritis in Madrid, Spain, 2022-2023.

In recent years, there has been an increase in Neisseria gonorrhoeae infections in Europe and Spain. Disseminated gonococcal infection is an uncommon clinical presentation that includes gonococcal arthritis. Improved antibiotic treatment has reduced the incidence of gonococcal arthritis. However, the increase in gonococcal infections may have increased the frequency of this clinical entity in recent times. We report five cases of gonococcal arthritis in patients in a tertiary-care hospital in the northern area of Madrid (Spain) from October 2022 to October 2023. Major cases occurred in male patients with unprotected sex and polyarticular symptoms requiring hospital admission and treatment with ceftriaxone and cefixime. The use of molecular techniques has allowed the detection of a greater number of culture-negative cases of gonococcal arthritis, as well as the detection of mutations associated with resistance to fluoroquinolone for switching to oral treatment.

Increase in congenital syphilis in a tertiary-care centre from Madrid, Spain: Back to the classics.

Globally, an increase in sexually transmitted diseases worldwide has been reported, including acquired Treponema pallidum infection. Congenital syphilis is the mother-to-child transmission syphilis infection, experiencing an increase in cases returning to incidence rates of years ago. We report three cases of symptomatic congenital syphilis occurring in a tertiary-care hospital in Madrid (Spain). Cases were diagnosed by serology and molecular biology methods. The use of molecular techniques in specimens such as skin lesion, subplacental exudate swabs, bronchoalveolar aspirate, or cerebrospinal fluid could favour the diagnosis of this clinical entity, especially in symptomatic newborns with systemic involvement.

Utility of culture and molecular methods using AllplexTM Bacterial Vaginosis Plus Assay (SeegeneⓇ) as a tool for endometriosis, infertility and recurrent pregnancy loss diagnosis.

Endometriosis, infertility, or recurrent pregnancy loss (RPL) are entities characterised by a decrease in Lactobacillus spp. and an increase in bacterial vaginosis-associated bacteria, (BVAV) according with 16S rRNA sequencing studies. However, the use of nucleic acid amplification tests (NAAT) as a tool for diagnosis algorithms is unknown. Seventy-four patients were included, with a median age of 36.5 years old (IQR: 34-39) including infertility (n=31), endometriosis (n=25), or RPL (n=18), for culturing and NAAT using the Allplex™ Bacterial Vaginosis Plus (ABVP) assay (SeegeneⓇ) with endometrial samples. The objective was determining the utility of ABVP assay for diagnosing the entities. Forty-six microorganisms were isolated from 31 out of 74 patients (41.9 %). Twenty-five endometrial samples (33.8 %) were positive for some targets included in the ABVP-assay, with median Ct value ∼37 (IQR: 31.3-37.1) and Qt value 1.43 Log10copies/reaction (IQR:1.1-2.6). For Lactobacillus species, sensitivity and specificity were 80 % and 84 %, respectively. Gardnerella vaginalis, 63.6 % and 95.7 %. No significant increase in BVAV was detected in any of the gynaecological entities. The ABVP and culture based algorithm did not show utility as a tool for endometriosis, infertility, or RPL diagnosis.

Sexually transmitted infections and clinical features in monkeypox (mpox) patients in Madrid, Spain.

Since the beginning of the monkeypox (mpox) virus outbreak in May 2022, there has been an increase in the number of cases worldwide in the setting of sexual transmission. We have tested by real-time PCR 187 mpox patients, of which 157 patients were screened for sexually transmitted infections (STI) in 245 samples. Thirty-six pathogens were detected in 30 patients: herpes simplex virus (HSV-I/II, 12/36, 33.3%), Neisseria gonorrhoeae (NG, 9/36, 25%), Chlamydia trachomatis (CT, 5/36, 13.8%), Chlamydia trachomatis-lymphogranuloma venereum (CT-LGV, 3/36, 8.3%), Treponema pallidum (TP, 4/36, 11.1%) and Mycoplasma genitalium (MG, 3/36, 8.3%). Screening of STI is recommended in mpox patients for the differential diagnosis of the main infections of sexual tract especially in patients with rectal involvement.