Field diagnosis and genotyping of chikungunya virus using a dried reverse transcription loop-mediated isothermal amplification (LAMP) assay and MinION sequencing.

Detection and sequencing of chikungunya virus (CHIKV) genome was performed using a combination of a modified reverse transcription loop-mediated isothermal amplification (RT-LAMP) method and a MinION sequencer. We developed the protocol for drying all the reagents for the RT-LAMP in a single reaction tube. Using this system, the CHIKV genome was effectively amplified under isothermal conditions, and used as a template for MinION sequencing with a laptop computer. Our in-house RT-LAMP method and MinION sequencing system were also validated with RNAs and serum samples from recent outbreaks of CHIKV patients in Brazil. The obtained sequence data confirmed the CHIKV outbreaks and identified the genotype. In summary, our established inexpensive on-site genome detection and sequencing system is applicable for both diagnosis of CHIKV infected patients and genotyping of the CHIKV virus in future outbreak in remote areas.

Morning blood pressure surge is associated with death in hypertensive patients

Background Morning blood pressure surge (MS), definedas the difference between the mean blood pressure (BP) 2 hafter waking up and the mean of the three lowest BP valuesduring sleep, has been correlated with increasedcardiovascular risk. We sought to evaluate its associationwith cardiovascular events and death.Methods We retrospectively analyzed data from 632hypertensive patients [median age 58 years (50–67 years),37% men] who underwent ambulatory BP monitoringbetween January 2005 and December 2006. Patients weredivided into two groups according to MS (≥41 mmHg and<41 mmHg), and mortality from any cause was retrievedafter a median time of 50 months (46–54 months).Results Patients with MS of 41 mmHg or higher were older,had a higher daytime systolic BP, as well as a highersystolic and diastolic dipping, and a lower night-timediastolic BP. During follow-up, there were 19 deaths and MSof 41 mmHg or higher was associated with a higher hazardfor death in the crude model [hazard ratio: 3.47 (95%confidence interval: 1.25–9.65)], as well as afteradjustments for age and the presence of diabetes [hazardratio: 3.35 (95% confidence interval: 1.18–9.49)].Conclusion An increased BP surge is associated withhigher hazard for death. Future studies specificallydesigned to evaluate the real impact of MS on outcomes, aswell as to define its optimal cutoff value, are required.