This paper reports the development of a one-step SYBR-Green I-based realtime RT-PCR assay for the detection and quantification of Chikungunya virus (CHIKV) in human, monkey and mosquito samples by targeting the E1 structural gene. A preliminary evaluation of this assay has been successfully completed using 71 samples, consisting of a panel of negative control sera, sera from healthy individuals, sera from patients with acute disease from which CHIKV had been isolated, as well as monkey sera and adult mosquito samples obtained during the chikungunya fever outbreak in Malaysia in 2008. The assay was found to be 100-fold more sensitive than the conventional RT-PCR with a detection limit of 4.12x10(0) RNA copies/μl. The specificity of the assay was tested against other related viruses such as Dengue (serotypes 1-4), Japanese encephalitis, Herpes Simplex, Parainfluenza, Sindbis, Ross River, Yellow fever and West Nile viruses. The sensitivity, specificity and efficiency of this assay were 100%, 100% and 96.8% respectively. This study on early diagnostics is of importance to all endemic countries, especially Malaysia, which has been facing increasingly frequent and bigger outbreaks due to this virus since 1999.

Objective To analyze the safety and effectiveness of robot-assisted laparoscopic partial nephrectomy(RLPN) for cT2 renal tumors in international multi-centers.Methods This study was conducted to collect information on surgical procedures performed by RLPN and robot assisted laparoscopic radical nephrectomy (RRN) in nineteen international urological centers from January 2012 to December 2017.RLPN were performed in 159 patients (118 males and 41 females),with the average age of (59.3 ± 13.2) years,body mass index(BMI) of (28.7 ± 5.4)kg/m2,preoperative GFR of (77.3 ± 22.1) ml/min.RRN were performed in 219 patients,with the average age of (62.0 ± 12.9) years,BMI of (28.7 ±6.1) kg/m2,preoperative GFR of (71.4 ± 20.3) ml/min.There was no statistical difference between the two groups in gender and BMI.The age of the patients in RLPN group was younger than that in RRN group,and the preoperative GFR was better.The patient's baseline demographics,perioperative data,tumor pathology,oncologic outcomes,and renal function (GFR) were recorded.Results All 378 cases underwent successful surgery.The operation time of RLPN was 150 min(65-353 min),which was shorter than that of RRN [180 min(85-361 min),P ＜ 0.001].The intra-operative blood loss of RLPN was more than that of RRN [150 ml (40-3 000 ml) vs.100 ml (10-1 100 ml),P ＜ 0.001].The incidence of intra-operative complications were not statistically different between the two groups [5.7％ (9/159) vs.3.2％ (7/219),P =0.240].The incidence of postoperative complications was higher in the RLPN group than that in RRN group [19.5％ (31/159) vs.10.5％ (23/219),P =0.014],but there was no significant difference in the incidence of complications of grade 3 or above [4.4％ (7/159) vs.2.3％ (5/219),P =0.246].The recurrence-free survival rate of RLPN group was higher than that of RRN group [91.4％ (117/128) vs.81.9％ (167/204),P =0.013],and RLPN group was more conducive to renal function protection (P ＜ 0.001).Conclusions RLPN for cT2 tumors can obtain effective tumor control rate and better renal function preservation.It could be an acceptable alternative for surgical management of cT2 tumors.