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ABSTRACT

Objectives: To identify and characterize respiratory viruses that infect children from daycare centers with symptoms of respiratory infection and to evaluate the association of clinical and epidemiological disease data with the identified virus. Methods:We conducted a study between 2003 and 2005 in 176 children with respiratory infection symptoms attending a municipal daycare center. Samples from nasopharyngeal secretion were tested by reverse transcriptase polymerase chain reaction and positive samples for picornavirus were sequenced.Results: All 782 collected samples were analyzed and 31.8% were positive for at least one of the studied respiratory viruses. Respiratory infections were characterized by the presence of mild symptoms of the upper respiratory tract, the most common of which were runny nose and cough. In the 2 years of study, most cases of infection occurred in autumn and winter, but respiratory viruses were detected throughout all the study period. Conclusions:Respiratory viruses and respiratory infections caused by them are part of the daily life of children attending daycare centers. Our results show the great impact that respiratory infections have on these children and suggest that more attention must be paid to viral pathogens. Resultados: Todas as 782 amostras coletadas foram analisadas, e 31,8% foram positivas para algum dos vírus respiratórios estudados. As infecções respiratórias foram caracterizadas pela presença de sintomas moderados do trato respiratório superior, sendo os mais comuns coriza e tosse. Em 2 anos de estudo, a maioria dos casos de infecção ocorreram no outono e no inverno, mas vírus respiratórios foram detectados ao longo de todo o período de estudo. J Pediatr (Rio J) Conclusões:Vírus respiratórios e as infecções respiratórias causadas por eles fazem parte do cotidiano de crianças que frequentam creches. Nossos resultados mostram o grande impacto gerado pelas infecções respiratórias nessas crianças e sugere que uma maior atenção deve ser dispensada aos patógenos virais.J Pediatr (Rio J). 2011;87(5):439-44: Vírus respiratórios, infecção respiratória aguda, crianças de creche.

One of the biggest challenges during the pandemic has been obtaining and maintaining critical material to conduct the increasing demand for molecular tests. Sometimes, the lack of suppliers and the global shortage of these reagents, a consequence of the high demand, make it difficult to detect and diagnose patients with suspected SARS-CoV-2 infection, negatively impacting the control of virus spread. Many alternatives have enabled the continuous processing of samples and have presented a decrease in time and cost. These measures thus allow broad testing of the population and should be ideal for controlling the disease. In this sense, we compared the SARS-CoV-2 molecular detection effectiveness by Real time RT-PCR using two different protocols for RNA extraction. The experiments were conducted in the National Institute of Health (INS) from Peru. We compared Ct values average (experimental triplicate) results from two different targets, a viral and internal control. All samples were extracted in parallel using a commercial kit and our alternative protocol–samples submitted to proteinase K treatment (3 μg/μL, 56°C for 10 minutes) followed by thermal shock (98°C for 5 minutes followed by 4°C for 2 minutes); the agreement between results was 100% in the samples tested. In addition, we compared the COVID-19 positivity between six epidemiological weeks: the initial two in that the Real time RT-PCR reactions were conducted using RNA extracted by commercial kit, followed by two other using RNA obtained by our kit-free method, and the last two using kit once again; they did not differ significantly. We concluded that our in-house method is an easy, fast, and cost-effective alternative method for extracting RNA and conducing molecular diagnosis of COVID-19.