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Zika virus (ZIKV) emergence in South America revealed the lack of knowledge regarding clinical manifestations in HIV-infected individuals.We described the clinical characteristics, laboratory manifestations, differential diagnosis, and outcome of ZIKV infection in a large, single-center cohort of HIV-infected patients.HIV-infected patients aged 18 years and older with clinical suspected arboviral disease from an ongoing cohort were followed from February 2015 through December 2015. Acute serum samples were tested for ZIKV, dengue virus (DENV), and chikungunya virus by real-time reverse transcriptase polymerase chain reaction, anti-DENV immunoglobulin (Ig)M/IgG, and syphilis assays; convalescent samples were tested for anti-DENV IgM/IgG; and urine samples were tested for ZIKV by real-time reverse transcriptase polymerase chain reaction. ZIKV disease was defined according to the Pan American Health Organization (PAHO) guidelines.Of 101 patients, ZIKV was confirmed in 43 cases and suspected in 34, and another diagnosis was assumed for 24 patients (dengue, secondary/latent syphilis, respiratory infections, human parvovirus B19, adverse drug reaction, musculoskeletal disorders, and acute gastroenteritis). ZIKV-confirmed and ZIKV-suspected patients reported similar signs and symptoms. Pruritic rash was the most common symptom, followed by myalgia, nonpurulent conjunctivitis, arthralgia, prostration, and headache. In the short-term follow-up [median 67.5 days (interquartile range: 32–104.5)], CD4 cell count (Z = −0.831, P = 0.406) and HIV viral load (Z = −0.447, P = 0.655) did not change significantly after ZIKV infection. There were no hospitalizations, complications, or deaths.Among HIV-infected patients with suspected arboviral disease, 42.6% were ZIKV-infected. CD4 cell counts and HIV viral load were not different after ZIKV infection. Differential diagnosis with other diseases and adverse drug reaction should be evaluated.