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Background: HIV-2 is a neglected virus despite estimates of 1-2 million people being infected worldwide. The virus is naturally resistant to some antiretrovirals used to treat HIV-1 and therapeutic options are limited for patients with HIV-2. Methods: In this retrospective observational study, we analysed all HIV-2-infected individuals treated with integrase strand transfer inhibitors (INSTIs) recorded in the Spanish HIV-2 cohort. Demographics, treatment modalities, laboratory values, quantitative HIV-2 RNA and CD4 counts as well as drug resistance were analysed. Results: From a total of 354 HIV-2-infected patients recruited by the Spanish HIV-2 cohort as of December 2017, INSTIs had been given to 44, in 18 as first-line therapy and in 26 after failing other antiretroviral regimens. After a median follow-up of 13 months of INSTI-based therapy, undetectable viraemia for HIV-2 was achieved in 89% of treatment-naive and in 65.4% of treatment-experienced patients. In parallel, CD4 gains were 82 and 126 cells/mm 3 , respectively. Treatment failure occurred in 15 patients, 2 being treatment-naive and 13 treatment-experienced. INSTI resistance changes were recognized in 12 patients: N155H (5), Q148H/R (3), Y143C/G (3) and R263K (1). Conclusions: Combinations based on INSTIs are effective and safe treatment options for HIV-2-infected individuals. However, resistance mutations to INSTIs are selected frequently in failing patients, reducing the already limited treatment options.

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Ovotesticular differences of sexual development (OT-DSD) are rare genetic variances defined by the coexistence of both testicular and ovarian tissues. Various molecular etiologies including SRY translocation or SOX9 pathogenic variants with different modes of inheritance have been associated with 46,XX OT-DSD. Here we describe a child diagnosed with SRY-negative 46,XX OT-DSD after completing a series of complex clinical genetic analyses, including chromosomal microarray, DSD gene panel (sequencing and deletion/duplication analysis), whole exome sequencing, and whole genome sequencing. Of these, only whole genome sequencing reported a pathogenic duplication in a non-coding region that contains the RevSex regulatory element, which modifies SOX9 expression and is associated with 46,XX OT-DSD and complete sex reversal. This is the first clinical RevSex duplication detected by clinical whole genome sequencing. We highlight the utility of whole genome sequencing in shortening the diagnostic odyssey and the importance of optimal counseling through a teambased multi-specialty approach for patients with DSDs.

BackgroundHuman papillomavirus (HPV) DNA testing plays a main role in the management of cervical cancer, however to improve the specificity in cervical screening, there is a need to develop and validate different approaches that can identify women at risk for progressive disease.Nowadays, mRNA expression of viral E6 and E7 HPV oncogenes stands up as a potential biomarker to improve cervical screening. We aimed to validate a method for RNA extraction, detect HPV mRNA expression and, assess the relationship between E6/E7 mRNA expression and pathology of patients’ lesions and progression.MethodsThis study included 50 specimens that had been previously genotyped as HPV16, 18, 31, 33 and/or 45. Cervical swabs were extracted with three different RNA extraction methods -Nuclisens manual extraction kit (bioMérieux), High Pure Viral RNA Kit (Roche) and RNeasy Plus Mini kit (Qiagen)-, and mRNA was detected with NucliSens EasyQ HPV version 1 test (bioMérieux) afterwards. Association of oncogene expression with pathology and lesion progression was analyzed for each extraction method.ResultsE6/E7 mRNA positivity rate was higher in samples analyzed with bioMérieux (62%), followed by Roche (24%) and Qiagen (6%). Women with lesions and lesion progression showed a higher prevalence of viral RNA expression than women that had not lesions or with lesion persistence. While bioMérieux revealed a higher sensitivity (77.27%), Roche presented a higher PPV (75%) and an increased specificity (89.28%).ConclusionsExtraction methods based on magnetic beads provided better RNA yield than those based in columns. Both Nuclisens manual extraction kit (bioMérieux) and High Pure Viral RNA Kit (Roche) seemed to be adequate for E6/E7 mRNA detection. However, none of them revealed both high sensitivity and specificity values. Further studies are needed to obtain and validate a standard gold method for RNA expression detection, to be included as part of the routine cervical screening program.

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HIV-2 is a retrovirus that mainly infects West Africans. In Europe, HIV-2 has been circulating since the 1980s, and more recent immigration has contributed to its spread. Excluding HIV-2 in all HIV seropositive individuals precludes misinterpretation of viral loads (VL) and antiretroviral (ART) choices. Surveillance registers enables tracking of epidemic spread and identification of major contributors, allowing the establishment of convenient preventive measures. The HIV-2 Spanish study group was founded in 1989. Since then, blood specimens from HIV-2 carriers have been collected. More than 40 Spanish hospitals are part of the group and provide clinical and epidemiological data. Records for each HIV-2 patient include country of origin, gender, age, transmission category, monitoring of HIV-2 VL, CD4 counts, drug resistance, and HIV-2 subtype. Up to December 2017, 354 HIV-2+ individuals were identified in Spain and incidence (15–20/year) has been stable within the last decade. At diagnosis, mean age was 44.6 years and 63 per cent were male. The majority were Africans (78%), whereas 16.5 per cent were native Spaniards. 78.2 per cent acquired HIV-2 by heterosexual contact. HIV-2 subtyping using the HIV2EU tool was performed in 126 subjects: 86 Africans and 27 native Spaniards. The subtype distribution was as follows: 108 (85.7%) HIV-2 subtype A and 18 (14.3%) B. Africans and Spaniards were mostly infected with subtype A (87.2% and 77.8%, respectively). HIV-2 subtype B was found in six native Spaniards (22.2%; 6/27), five patients from Equatorial Guinea (71.4%; 5/7), four from Senegal (18.1%; 4/22), two from Ivory Coast (100%; 2/2), and one from Burkina Faso (50%; 1/2). Using phylogenetic analyses, two clusters were identified among homosexual Spanish men (subtype A: 8 men and subtype B: 2 men with viral isolates related to Malian and Senegalese isolates). Before starting ART, CD4 count mean values for subtype A and B were 378 and 357, respectively. Corresponding VL values were 2.63 and 2.32 HIV-2 RNA log copies/ml, respectively.