RT-LAMP assay for ultra-sensitive detection of SARS-CoV-2 in saliva and VTM clinical samples

The COVID-19 pandemic has underscored the shortcomings in the deployment of state-of-the-art diagnostic platforms. Although several PCR-based techniques have been rapidly developed to meet the growing testing needs, such techniques often need samples collected through a swab, the use of RNA extraction kits, and expensive thermocyclers in order to successfully perform the test. Isothermal amplification-based approaches have also been recently demonstrated for rapid SARS-CoV-2 detection by minimizing sample preparation while also reducing the instrumentation and reaction complexity. There are limited reports of saliva as the sample source and some of these indicate inferior sensitivity when comparing RT-LAMP with PCR-based techniques. In this paper, we demonstrate an improved sensitivity assay to test saliva using a 2-step RT-LAMP assay, where a short 10-minute RT step is performed with only B3 and BIP primers before the final reaction. We show that while the 1-step RT-LAMP demonstrate satisfactory results, the optimized 2-step approach allows for single molecule sensitivity per reaction and performs significantly better than the 1-step RT-LAMP and conventional 2-step RT-LAMP approaches with all primers included in the RT Step. Importantly, we demonstrate RNA extraction-free RT-LAMP based assays for detection of SARS-CoV-2 from VTM and saliva clinical samples.

Going home: what NICU nurses need to know about home care.

Pediatric home health care enables patients to be at home with their families in settings that bring them joy, comfort, and the security we all feel when we are at home. There is also a feeling, no matter how small, that the parents have some control over what is happening to their child. Infants with multiple needs require in-depth discharge planning. There are the physical and health concerns of the preterm infant and the potential complications that he could still develop. Parent teaching is vital for the successful transition from hopital to home. When the neonatal discharge nurse is aware of what difficulties the parents and the patient might face at home, her teaching can be tailored to meer the specific needs of these vulnerable, complicated infants. This article discusses the discharge planning process, which begins upon admission to the neonatal intensive care unit, as well as common problems encountered by many premature infants discharged home.