Potato aucuba mosaic virus (PAMV) ELISA Kit

Potato viruses including Potato aucuba mosaic virus (PAMV), Potato leafroll virus (PLRV), Potato virus M (PVM), Potato virus S (PVS) and Potato virus X (PVX) are well known viruses infecting potato and can cause serious economic losses. In this study, a multiplex reverse transcription polymerase chain reaction (RT-PCR) assay was developed and standardized for simultaneous detection of PAMV, PLRV, PVM, PVS and PVX with elongation factor 1-alpha (ef1 alpha) as plant internal control. At least three primer pairs were designed within the conserved regions of genomic sequences of five potato viruses and ef1 alpha of potato for single and/or multiplex RT-PCR. The reaction conditions were initially optimized by selecting primer pairs and standardized the individual RT-PCR. Then, one pair of primer for each virus including ef1 alpha was selected for multiplex RT-PCR producing distinct fragments of 305, 452, 408, 363, 565, 803 bp, representing PAMV, PLRV, PVM, PVS, PVX and ef1 alpha, respectively. All the primers were designed based on specificity and compatibility and the reliability was confirmed by cloning and sequencing followed by BLAST analysis. The sensitivity of single as well as multiplex RT-PCR was evaluated. The assay could detect all five viruses including ef1 alpha even at total RNA dilutions of 10(-3) and total cDNA dilution of 10(-7). The multiplex RT-PCR assay was then used to test virus infections from field samples of potato (leaves and tubers) collected from different states of India along with plant internal control. This multiplex RT-PCR was equally enough in detecting mixed infection of five viruses in foliage as well as tubers. The results indicate the robustness and reliability of the assay for virus indexing of mother stocks for five viruses simultaneously in potato seed production and will be useful for seed certification programme, breeding and epidemiological studies.

Background: The aim of our study was to evaluate the prognostic value of Charlson's weighted index of comorbidities (WIC) in patients with prolonged acute mechanical ventilation (PAMV, ventilator care >= 96 hours). Methods: We retrospectively enrolled 299 Korean PAMV patients who were admitted in a medical intensive care unit (ICU) of a university-affiliated tertiary care hospital between 2008 and 2013. Survivors were defined as patients who survived for 60 days after ICU admission. Results: The patients' mean age was 65.1 +/- 14.1 years and 70.6% were male. The mean ICU and hospital length of stay was 21.9 +/- 19.7 and 39.4 +/- 39.1 days, respectively. In addition, the 60-day mortality rate after ICU admission was 35.5%. The mean WIC was 2.3 +/- 1.8, with significant differences between nonsurvivors and survivors (2.7 +/- 2.1 vs. 2.1 +/- 1.7, p<0.05). The area under the curve of receiver-operating-characteristics curve for WIC was 0.593 (95% confidence interval [CI], 0.523-0.661; p<0.05). Based on Kaplan-Meier curves of 60-day survival, WIC >= 5 had statistically lower survival than WIC <5 (log-rank test, p<0.05). In a multivariate Cox proportional hazard model, WIC >= 5 was associated with poor prognosis (hazard ratio, 1.901; 95% CI, 1.140-3.171; p<0.05). The mortality rate of patients with WIC >= 5 was 54.2%. Conclusion: Our study showed a WIC score >= 5 might be helpful in predicting 60-day mortality in PAMV patients.