Ten-minute direct detection of Zika virus in serum samples by RT-LAMP. 2019

Paulo Felipe Neves Estrela, and Geovana de Melo Mendes, and Kézia Gomes de Oliveira, and Alexandre Melo Bailão, and Célia Maria de Almeida Soares, and Nilson Antônio Assunção, and Gabriela Rodrigues Mendes Duarte
Instituto de Química, Universidade Federal de Goiás, Goiânia, GO, 74690-900, Brazil.

Zika virus (ZIKV) is a current threat to global health. In most of cases, ZIKV infection has no symptoms; however in some cases, ZIKV can cause paralysis (Guillain-Barré syndrome), and in pregnant women, it can cause birth defects in infants. Rapid and accurate diagnosis can help improve disease control as well as being vital to prenatal care for women living in endemic areas. Molecular diagnostics based on isothermal amplification techniques are an excellent alternative to conventional methods of DNA amplification, such as PCR. Here, we develop and optimized a rapid and sensitive method for direct detection of ZIKV in Serum samples based on RT-LAMP and visual detection. The reaction was thermally controlled with a thermoblock for 10 min at 72 °C. The results show that the use of the Bst 3.0 enzyme and an adequate optimization can further reduce the time needed for the RT-LAMP reaction to detect ZIKV. Our results demonstrate that it is possible to detect ZIKV through RT-LAMP directly from a Serum sample, without prior RNA extraction. As little as 10-3 copies of RNA in a 10 μL reaction (20 zepto-molar) was detected by RT-LAMP from a panel of 51 Serum samples (16 samples from pregnant women and 35 samples from newborns infected with ZIKV during pregnancy). The RT-LAMP has proven to be a valuable tool for molecular diagnosis of Zika, presenting a great potential for point-of-care applications, especially in developing countries.

UI MeSH Term Description Entries
D007231 Infant, Newborn An infant during the first 28 days after birth. Neonate,Newborns,Infants, Newborn,Neonates,Newborn,Newborn Infant,Newborn Infants
D011247 Pregnancy The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH. Gestation,Pregnancies
D005260 Female Females
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000071243 Zika Virus Infection A viral disease transmitted by the bite of AEDES mosquitoes infected with ZIKA VIRUS. Its mild DENGUE-like symptoms include fever, rash, headaches and ARTHRALGIA. The viral infection during pregnancy, in rare cases, is associated with congenital brain and ocular abnormalities, called Congenital Zika Syndrome, including MICROCEPHALY and may also lead to GUILLAIN-BARRE SYNDROME. Congenital Zika Syndrome,Congenital Zika Virus Infection,Fever, Zika,ZikV Infection,Zika Fever,Zika Virus Disease,Disease, Zika Virus,Infection, ZikV,Infection, Zika Virus,Virus Disease, Zika,Virus Infection, Zika
D000071244 Zika Virus An arbovirus in the FLAVIVIRUS genus of the family FLAVIVIRIDAE. Originally isolated in the Zika Forest of UGANDA it has been introduced to Asia and the Americas. Virus, Zika,ZikV
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D019095 Point-of-Care Systems Laboratory and other services provided to patients at the bedside. These include diagnostic and laboratory testing using automated information entry. Bedside Computing,Point of Care Technology,Bedside Technology,Point-of-Care,Bedside Technologies,Computing, Bedside,Point of Care,Point of Care Systems,Point-of-Care System,Systems, Point-of-Care,Technologies, Bedside,Technology, Bedside

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