The molecular identification of ZIKV in the amniotic fluid and cerebrospinal fluid was done by using a SuperScript III Platinum One-Step RT-PCR system (Invitrogen, Carlsbad, CA, United States) run by the Colombian Government National Institutes of Health using the primers ZIKV 1087, ZIKV 1163, and ZIKV 1108-FAM, followed by the Lanciotti protocol (Lanciotti et al., 2008 (link)). For hybridization and extension, an ABI 7500 Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, United States) was operated (Pacheco et al., 2016 (link)).
Samples were obtained from the mother and the newborn and those from viral isolation trials for ZIKV RNA, using a TaqMan RT-PCR assay and quantitative RT-PCR for ZIKV following the CDC protocol (Centers for Disease Control and Prevention, 2019 ). Standard methods were used to determine the levels of ZIKV IgM, IgG, and neutralizing antibody titers. Additionally, we tested for other flaviviruses and arthropod-borne viruses. In addition, the antibody levels of infectious diseases that might produce congenital abnormalities, such as toxoplasma, cytomegalovirus, syphilis, herpes, rubella, hepatitis, parvovirus B19, and HIV, were tested in the mother and the newborn (TORCH test), as is described previously in the literature (Candelo et al., 2019 (link)).
Samples were obtained from the mother and the newborn and those from viral isolation trials for ZIKV RNA, using a TaqMan RT-PCR assay and quantitative RT-PCR for ZIKV following the CDC protocol (Centers for Disease Control and Prevention, 2019 ). Standard methods were used to determine the levels of ZIKV IgM, IgG, and neutralizing antibody titers. Additionally, we tested for other flaviviruses and arthropod-borne viruses. In addition, the antibody levels of infectious diseases that might produce congenital abnormalities, such as toxoplasma, cytomegalovirus, syphilis, herpes, rubella, hepatitis, parvovirus B19, and HIV, were tested in the mother and the newborn (TORCH test), as is described previously in the literature (Candelo et al., 2019 (link)).