1.Effects of different intensity interval exercise of 6 weeks on body composition of obese rats.
Shu-Qiang ZHAO ; Hong-Ju SHI ; Ning-Ning ZHENG
Chinese Journal of Applied Physiology 2019;35(4):326-330
		                        		
		                        			OBJECTIVE:
		                        			To investigate the effects of different intensity intermittent exercise on the body function of obese rats, and to provide basis for the prevention and treatment of obesity.
		                        		
		                        			METHODS:
		                        			Eighty SD rats were randomly divided into normal diet group (n=20) and high-fat diet group (n=60). After adaptive feeding for 8 weeks, 8 normal diet rats and 32 high-fat obese rats were selected for follow-up experiments. The experimental rats were randomly divided into 5 groups (n=8): control diet-sedentary (CS),with ordinary feed and without any exercise; high diet-sedentary (HS), with high-fat feed feeding and without any exercise; high diet-continual exercise(HC), 60 min/day,5 days/week with 6 weeks; high diet-long time-low frequency interval exercise(HLL), 30 min/time,twice/day (intermittent 6 h), 5 days/week with 6 weeks; high diet-short time-high frequency interval exercise(HSH), 20 min/ time, 3 times/d (intermittent 3 h), 5 days/week with 6 weeks. The training intensity of rats in each exercise group was 25 m/min. After 6 weeks, rats in each groups were weighed, and resting metabolic rate(RMR), fasting blood glucose(FBG), triglyceride(TG) and other biochemical indexes were detected, and fat and muscle weight were measured.
		                        		
		                        			RESULTS:
		                        			Before experiment, there were no significant differences in RMR, FBG and TG in each groups(P>0.05).The body weight of HSH, HLL, HC and HS groups was higher than that of CS group (P<0.05). After the experiment, RMR of the HSH,HLL and HC groups was significantly higher than that of HS and CS groups (P<0.05), but without significant difference among the HSH,HLL and HC groups (P>0.05).The body weight of HSH, HLL and HC groups was significantly lower than that of HS group (P<0.05), but the three groups was not significant (P>0.05); perirenal fat(PF), idymis fat(EF), perirenal fat/weight(PF/W) and epididymis fat/weight(EF/W) of HSH, HLL, HC group were significantly lower than those of HS group (P<0.01), while there was no statistical difference among the three groups (P>0.05). There was no significant difference in gastrocnemius(GM) and quadricep(QF) of each group (P>0.05), gastrocnemius/weight(GM/W) and quadriceps/weight (QF/W) in HSH,HLL and HC groups were higher than those of HS group (P<0.05),while there was no significant difference among HSH,HLL and HC groups (P>0.05);FEB,TG of HSH,HLL,HC group were lower than those of CS and HS group (P<0.05),but the difference with HS group was more significant (P<0.01),there was no significant difference among training groups(P>0.05).
		                        		
		                        			CONCLUSION
		                        			6 weeks of intermittent exercise of different intensity had a good intervention effect on the body composition of obese rats, and high diet-short time-high frequency interval exercise (HSH) may be more effective.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Body Composition
		                        			;
		                        		
		                        			Body Weight
		                        			;
		                        		
		                        			Diet, High-Fat
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Obesity
		                        			;
		                        		
		                        			therapy
		                        			;
		                        		
		                        			Physical Conditioning, Animal
		                        			;
		                        		
		                        			Random Allocation
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			
		                        		
		                        	
2.Bioinformation analysis of chorismate synthase in Baphicacantus cusia and other 78 species of plants.
Jian YU ; Qi YE ; Shu-Ju NING ; Qing LI ; He-Xin TAN ; Jing-Xian FENG ; Rui-Bing CHEN ; Xiao-Li MA ; Pei-Min GONG ; Xuan-Xuan ZHAO ; Lei ZHANG ; Dao-Zhi WEI
China Journal of Chinese Materia Medica 2018;43(4):721-730
		                        		
		                        			
		                        			Chorismate synthase(CS, EC:4.2.3.5) catalyses 5-enolpyruvy-shikimate-3-phosphate to form chorismate, which is the essential enzyme for chorismate biosynthesis in organisms. The amino acid sequences of CS from 79 species of higher plants were reported in GenBank at present. 125 amino acid sequences of CS from Baphicacanthus cusia and other 78 species of plants were predicted and analyzed by using various bioinformatics software, including the composition of amino acid sequences, signal peptide, leader peptide, hydrophobic/hydrophilic, transmembrane structure, coiled-coil domain, protein secondary structure, tertiary structure and functional domains. The phylogenetic tree of CS protein family was constructed and divided into eight groups by phylogenetic analysis. The homology comparison indicated that B. cusia shared a high homology with several plants such as Sesamum indicum, Nicotiana tabacum, Solanum tuberosum and so on. The open reading frame(ORF) of all samples is about 1 300 bp, the molecular weight is about 50 kDa, the isoelectric point(pI) is 5.0-8.0 which illustrated that CS protein is slightly basic. The ORF of CS we cloned in B. cusia is 1 326 bp, the amino acid residues are 442, the molecular weight is 47 kDa and pI is 8.11. The CS in B.cusia showed obvious hydrophobicity area and hydrophilicity area, no signal peptide, and may exists transmembrane structure areas. The main secondary structures of CS protein are random coil and Alpha helix, also contain three main structural domains which are an active structural domain, a PLN02754 conserved domain and a FMN binding site. The acquired information in this study would provide certain scientific basis for further study on structure-activity relationship and structure modification of CS in plants in the future.
		                        		
		                        		
		                        		
		                        			Acanthaceae
		                        			;
		                        		
		                        			enzymology
		                        			;
		                        		
		                        			Amino Acid Sequence
		                        			;
		                        		
		                        			Computational Biology
		                        			;
		                        		
		                        			Phosphorus-Oxygen Lyases
		                        			;
		                        		
		                        			chemistry
		                        			;
		                        		
		                        			Phylogeny
		                        			;
		                        		
		                        			Plant Proteins
		                        			;
		                        		
		                        			chemistry
		                        			;
		                        		
		                        			Protein Structure, Secondary
		                        			
		                        		
		                        	
3.Effect of Kuanxiong Aerosol () on Patients with Angina Pectoris: A Non-inferiority Multi-center Randomized Controlled Trial.
Qiao-Ning YANG ; Rui-Na BAI ; Guo-Ju DONG ; Chang-Jiang GE ; Jing-Min ZHOU ; Li HUANG ; Yan HE ; Jun WANG ; Ai-Hua REN ; Zhan-Quan HUANG ; Guang-Li ZHU ; Shu LU ; Shang-Quan XIONG ; Shao-Xiang XIAN ; Zhi-Jun ZHU ; Da-Zhuo SHI ; Shu-Zheng LU ; Li-Zhi LI ; Ke-Ji CHEN
Chinese journal of integrative medicine 2018;24(5):336-342
OBJECTIVETo evaluate the effect and safety of Kuanxiong Aerosol (, KA) on patients with angina pectoris.
METHODSBlock randomization was performed to randomly allocate 750 patients into KA (376 cases) and control groups (374 cases). During an angina attack, the KA group received 3 consecutive sublingual sprays of KA (0.6 mL per spray). The control group received 1 sublingual nitroglycerin tablet (NT, 0.5 mg/tablet). Log-rank tests and Kaplan-Meier estimations were used to estimate the angina remission rates at 6 time-points after treatment (1, 2, 3, 4, 5, and >5 min). Logistic regression analysis was performed to observe the factors inflfluencing the rate of effective angina remission, and the remission rates and incidences of adverse reactions were compared for different Canadian Cardiovascular Society (CCS) classes of angina.
RESULTSThe 5-min remission rates in the KA and control groups were not signifificantly different (94.41% vs. 90.64%, P>0.05). The angina CCS class signifificantly inflfluenced the rate of remission (95% confidence interval = 0.483-0.740, P<0.01). In the CCS subgroup analysis, the 3-and 5-min remission rates for KA and NT were similar in the CCSII and III subgroups (P>0.05), while they were signifificantly better for KA in the CCSI and II subgroups (P<0.05 or P<0.01). Furthermore, the incidence of adverse reactions was signifificantly lower in the KA group than in the control group for the CCSII and III subgroups (9.29% vs. 26.22%, 10.13% vs. 20.88%, P<0.05 or P<0.01).
CONCLUSIONSKA is not inferior to NT in the remission of angina. Furthermore, in CCSII and III patients, KA is superior to NT, with a lower incidence of adverse reactions. (Registration No. ChiCTRIPR-15007204).
Aerosols ; adverse effects ; therapeutic use ; Angina Pectoris ; drug therapy ; Case-Control Studies ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Kaplan-Meier Estimate ; Logistic Models ; Male ; Middle Aged ; Remission Induction ; Treatment Outcome
4.Effect of atorvastatin on the expression of fractalkine in the circulation of patients with acute coronary syndrome
Kang YAO ; Hai-Bo LIU ; Shu-Ning ZHANG ; Xiao-Wu HONG ; Hao LU ; Ju-Ying QIAN ; Jun-Bo GE
Chinese Journal of Clinical Medicine 2016;23(3):269-272
		                        		
		                        			
		                        			Objective:To observe the effect of atorvastatin on the expression of soluble fractalkine in the circulation of patients with acute coronary syndrome .Methods:40 patients with acute coronary syndrome (ACS ) confirmed by coronary angiography were selected ,including 20 patients with acute myocardial infarction (AMI group) and20 patients with unstable angina pectoris (UAP group) .Another 20 hyperlipidemia patients with negative coronary angiography were selected as the control group .All three groups were given atorvastatin 20 mg per time ,one time per day for 3 months .Fractalkine levels in elbow vein blood were detected before and after treatment by ELISA method . CX3CR1 expression in peripheral blood mononuclear cells (PBMC) was examined by flow cytometry .Results:On admission ,fractalkine levels in the circulation of AMI group ([1047 ± 348 .9] pg/mL) and UAP Group ([777 .2 ± 324 .4] pg/mL) were significantly higher than that of the control group ([485 .1 ± 26 .09] pg /mL , P< 0 .001 , P= 0 .004);CX3CR1 expression rates of CD14+ monocytes in the circulation of AMI group ([38 .8 ± 15 .4]% ) and UAP group ([29 .2 ± 11 .9]% ) were significantly higher than that of the control group ([15 .2 ± 7 .9]% , P= 0 .009 , P= 0 .03) .After 3 months of atorvastatin treatment ,AMI ([740 .1 ± 269 .9] pg/mL) and UAP ([523 .8 ± 188 .5]pg/mL) fractalkine levels were significantly lower than before treatment (P= 0 .005 , P=0 .01 ) .Conclusions: Atorvastatin can significantly reduce fractalkine expression in the circulation of acute coronary syndrome patients ,and then achievethe immune effect of anti-atherosclerosis inflammation .
		                        		
		                        		
		                        		
		                        	
5.Does Early Postsurgical Temozolomide Plus Concomitant Radiochemotherapy Regimen Have Any Benefit in Newly-diagnosed Glioblastoma Patients? A Multi-center, Randomized, Parallel, Open-label, Phase II Clinical Trial.
Ying MAO ; Yu YAO ; Li-Wei ZHANG ; Yi-Cheng LU ; Zhong-Ping CHEN ; Jian-Min ZHANG ; Song-Tao QI ; Chao YOU ; Ren-Zhi WANG ; Shu-Yuan YANG ; Xiang ZHANG ; Ji-Sheng WANG ; Ju-Xiang CHEN ; Qun-Ying YANG ; Hong SHEN ; Zhi-Yong LI ; Xiang WANG ; Wen-Bin MA ; Xue-Jun YANG ; Hai-Ning ZHEN ; Liang-Fu ZHOU
Chinese Medical Journal 2015;128(20):2751-2758
BACKGROUNDThe radiochemotherapy regimen concomitantly employing temozolomide (TMZ) chemotherapy and radiotherapy (RT) 4 weeks after surgery, followed by 6 cycles of TMZ is a common treatment for glioblastoma (GBM). However, its median overall survival (OS) is only 14.6 months. This study was to explore the effectiveness and safety of early TMZ chemotherapy between surgery and chemoradiotherapy plus the standard concomitant radiochemotherapy regimen.
METHODSA randomized, parallel group, open-label study of 99 newly diagnosed GBM patients was conducted at 10 independent Chinese neurosurgical departments from June 2008 to June 2012. Patients were treated with concomitant radiochemotherapy regimen plus early postsurgical temozolomide (early TMZ group) or standard concomitant radiochemotherapy regimen (control group). Overall response was assessed based on objective tumor assessments, administration of corticosteroid and neurological status test. Hematological, biochemical, laboratory, adverse event (AE), and neurological condition were measured for 24 months of follow-up. The primary efficacy endpoint of this study was overall survival (OS). The secondary endpoint was progression free survival (PFS).
RESULTSThe median OS time in the early TMZ group was 17.6 months, compared with 13.2 months in the control group (log-rank test P = 0.021). In addition, the OS rate in the early TMZ group was higher at 6, 12, and 18 months than in the control group, respectively (P < 0.05). The median PFS time was 8.7 months in the early TMZ group and 10.4 months in the control group (log-rank test P = 0.695). AEs occurred in 29 (55.8%) and 31(73.8%) patients respectively in early and control groups, including nausea (15.4% vs. 33.3%), vomiting (7.7% vs. 28.6%), fever (7.7% vs. 11.9%), and headache (3.8% vs. 23.8%). Only 30.8% and 33.3% were drug-related, respectively.
CONCLUSIONSAddition of TMZ chemotherapy in the early break of the standard concomitant radiochemotherapy regimen was well tolerated and significantly improved the OS of the GBM patients, compared with standard concomitant radiochemotherapy regimen. However, a larger randomized trial is warranted to verify these results.
Adult ; Aged ; Antineoplastic Agents, Alkylating ; therapeutic use ; Chemoradiotherapy ; methods ; Dacarbazine ; analogs & derivatives ; therapeutic use ; Glioblastoma ; drug therapy ; radiotherapy ; Humans ; Middle Aged ; Treatment Outcome ; Young Adult
6.A Matched Comparison Study of Uniportal Versus Triportal Thoracoscopic Lobectomy and Sublobectomy for Early-stage Nonsmall Cell Lung Cancer.
Ju-Wei MU ; Shu-Geng GAO ; Qi XUE ; Jun ZHAO ; Ning LI ; Kun YANG ; Kai SU ; Zhu-Yang YUAN ; Jie HE
Chinese Medical Journal 2015;128(20):2731-2735
BACKGROUNDBoth uniportal and triportal thoracoscopic lobectomy and sublobectomy are feasible for early-stage non-small cell lung cancer (NSCLC). The aim of this study was to compare the perioperative outcomes of uniportal and triportal thoracoscopic lobectomy and sublobectomy for early-stage NSCLC.
METHODSA total of 405 patients with lung lesions underwent thoracoscopic lobectomy or sublobectomy through a uniportal or triportal procedure in approximately 7-month period (From November 2014 to May 2015). A propensity-matched analysis, incorporating preoperative variables, was used to compare the short-term outcomes of patients who received uniportal or triportal thoracoscopic lobectomy and sublobectomy.
RESULTSFifty-eight patients underwent uniportal and 347 patients underwent triportal pulmonary resection. The conversion rate for uniportal and triportal procedure was 3.4% (2/58) and 2.3% (8/347), respectively. The complication rate for uniportal and triportal procedure was 10.3% and 9.5%, respectively. There was no perioperative death in either group. Most patients had early-stage NSCLC in both groups (uniportal: 45/47, 96%; triportal: 313/343, 91%). Propensity score-matching analysis demonstrated no significant differences in operation time, intraoperative blood loss, numbers of dissected lymph nodes, number of stations of lymph node dissected, duration of chest tube, and complication rate between uniportal and triportal group for early-stage NSCLC. However, the duration of postoperative hospitalization was longer in the uniportal group (6.83 ± 4.17 vs. 5.42 ± 1.86 d, P = 0.036) compared with the triportal group.
CONCLUSIONSUniportal thoracoscopic lobectomy and sublobectomy is safe and feasible, with comparable short-term outcomes with triportal thoracoscopic pulmonary resection. Uniportal lobectomy and sublobectomy lead to similar cure rate as triportal lobectomy and sublobectomy for early NSCLC.
Adult ; Aged ; Blood Loss, Surgical ; Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Female ; Humans ; Length of Stay ; Lung ; pathology ; surgery ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Operative Time ; Pneumonectomy ; adverse effects ; methods ; Prospective Studies ; Thoracic Surgery, Video-Assisted ; adverse effects ; methods ; Treatment Outcome
7.Virological characterization of influenza A(H3N2) virus in Mainland China during 2011-2012.
Wei-Juan HUANG ; Yan-Hui CHENG ; Xi-Yan LI ; Xiang ZHAO ; Jun-Feng GUO ; Zhao WANG ; Min-Ju TAN ; Ming LI ; Hong-Tao SUI ; He-Jiang WEI ; Yao-Yao CHEN ; Ning XIAO ; Yu LAN ; Da-Yan WANG ; Yue-Long SHU
Chinese Journal of Virology 2013;29(3):258-264
		                        		
		                        			
		                        			To study the prevalence and variation of influenza A(H3N2) viruses, the antigenic and genetic characteristics of influenza A(H3N2) viruses circulating in Mainland China during April 2011 to March 2012 were analyzed. The results showed that influenza A(H3N2) viruses increased gradually since 2012 and became the dominant strain since March. The viruses were antigenically closely related to the vaccine strain A/PER/16/09 (87.2%) and the representative virus A/FJ/196/09 (76.0%) in Mainland China. The genetic characteristics analysis results showed that recently isolated viruses belonged to the Vic/208 clade, and most of the low reaction strains also fell into the same clade. Crystal structure analysis of HA protein found that, compared with the vaccine strain A/PER/16/09, the recently isolated viruses had amino acid substitutions in the antigenic site A, B and C areas, in addition to gaining potential glycosylation sites at the amino acid position of 45 of HA and 367 of NA. Although the majority of circulating influenza A (H3N2) viruses in 2011-2012 season in Mainland China were antigeniclly matched by current influenza vaccine strain and the selected representative viruses, low reaction strains have increased since 2012, therefore it is necessary to strengthen the surveillance on the variation of influenza virus and to provide solid information for the vaccine strain selection.
		                        		
		                        		
		                        		
		                        			Amino Acid Sequence
		                        			;
		                        		
		                        			China
		                        			;
		                        		
		                        			epidemiology
		                        			;
		                        		
		                        			Hemagglutinin Glycoproteins, Influenza Virus
		                        			;
		                        		
		                        			chemistry
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		                        			genetics
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		                        			Humans
		                        			;
		                        		
		                        			Influenza A Virus, H3N2 Subtype
		                        			;
		                        		
		                        			classification
		                        			;
		                        		
		                        			genetics
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		                        			isolation & purification
		                        			;
		                        		
		                        			physiology
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		                        			Influenza, Human
		                        			;
		                        		
		                        			epidemiology
		                        			;
		                        		
		                        			virology
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		                        			Models, Molecular
		                        			;
		                        		
		                        			Molecular Sequence Data
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		                        			Phylogeny
		                        			
		                        		
		                        	
8.Emerged Pdm09 influenza virus increased purifying selection of seasonal H1N1 influenza virus.
Yu LAN ; Wei-Juan HUANG ; Hong-Tao SUI ; Xi-Yan LI ; Xiang ZHAO ; Ming LI ; Yao-Yao CHEN ; Jun-Feng GUO ; Yan-Hui CHENG ; Min-Ju TAN ; Zhao WANG ; Ning XIAO ; He-Jiang WEI ; Da-Yan WANG ; Yue-Long SHU
Chinese Journal of Virology 2013;29(2):143-147
		                        		
		                        			
		                        			Pdm09 virus outbreak occurred in Mainland China in May 2009, a few months later, the prevalence of seasonal H1N1(sH1N1) influenza virus that already circulated in human for tens of years began to decline and disappeared afterwards. To identify the reason for the rapid decline of sH1N1 in mainland China, we sequenced the HA1 of sH1N1 during 2006-2011, and then analyzed the selective pressure in different phases. Our results showed before Pdm09 outbreak, the omega value was 0. 36 while after Pdm09 outbreak the omega value was 0. 28 and significant difference (t test, P<0. 05) was identified. We concluded that sH1N1 obtained stronger purifying selection after Pdm09 outbreak in China. This might one of the major reasons causing the disappearance of sH1N1 in human.
		                        		
		                        		
		                        		
		                        			China
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Influenza A Virus, H1N1 Subtype
		                        			;
		                        		
		                        			classification
		                        			;
		                        		
		                        			genetics
		                        			;
		                        		
		                        			isolation & purification
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		                        			Influenza, Human
		                        			;
		                        		
		                        			virology
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		                        			Phylogeny
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		                        			Seasons
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		                        			Selection, Genetic
		                        			
		                        		
		                        	
9.Virological characterization of influenza B virus in mainland China during 2011-2012.
Wei-Juan HUANG ; Min-Ju TAN ; Yu LAN ; Yan-Hui CHENG ; Zhao WANG ; Xi-Yan LI ; Jun-Feng GUO ; He-Jiang WEI ; Yao-Yao CHEN ; Ning XIAO ; Bin LIU ; Hong-Tao SUI ; Xiang ZHAO ; Da-Yan WANG ; Yue-Long SHU
Chinese Journal of Virology 2013;29(1):32-38
		                        		
		                        			
		                        			In order to understand the prevalence and variation of influenza B viruses, the antigenic and genetic characteristics of influenza B viruses circulating in Mainland China during April, 2011 to March, 2012 were analyzed. The results showed the B Victoria lineage viruses were much more prevalent than B Yamagata lineage during this period, phylogenetic analysis showed vast majority of Victoria lineage viruses belong to genetic group 1, intra-clade reassortant between HA1 and NA gene was identified in a minor proportion of the viruses. 72.8% of the B/Victoria-lineage viruses were antigenically closely related to the vaccine strain B/Brisbane/60/2008. B Yamagata component was not included in the trivalent influenza vaccine in China during the study period, however vast majority of B Yamagata lineage viruses were antigenically and genetically closely related to the representative virus B/Hubei-Wujiagang/158/2009(97.8%) and B/Sichuan-Anyue/139/2011(85.2%) in China, reassortant between HA1 and NA was not identified in B Yamagata lineage viruses. Overall, the predominant circulating influenza B viruses in 2011-2012 season in China were matched by current influenza vaccine and the selected representative viruses were proved to represent the antigenic and genetic characteristics of the circulating viruses.
		                        		
		                        		
		                        		
		                        			China
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Influenza B virus
		                        			;
		                        		
		                        			classification
		                        			;
		                        		
		                        			genetics
		                        			;
		                        		
		                        			immunology
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		                        			Influenza Vaccines
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		                        			genetics
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		                        			immunology
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		                        			Phylogeny
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		                        			Time Factors
		                        			
		                        		
		                        	
10.Development of a real-time reverse transcriptase PCR assay for detection of E119V amino acid change in neuraminidase of influenza A (H3N2) using the TaqMan-MGB probe.
Xiang ZHAO ; Wei-juan HUANG ; He-jiang WEI ; Zhao WANG ; Xi-yan LI ; Yan-hui CHENG ; Min-ju TAN ; Ning XIAO ; Yu LAN ; Jun-feng GUO ; Hong-tao SUI ; Wen-fei ZHU ; Dong-dong DU ; Da-yan WANG ; Yue-long SHU
Chinese Journal of Preventive Medicine 2013;47(5):448-451
OBJECTIVETo develop a rapid duplex Real-time reverse transcription PCR (rRT-PCR) method to detect E119V mutation on neuraminidase (NA) of influenza A(H3N2) subtype with drug resistance to oseltamivir.
METHODSTwenty-six NA genes of influenza A(H3N2) virus between 2000 and 2012 in GenBank database were selected as the target genes, and specific TaqMan-MGB probe was designed to target the E119V amino acid change in neuraminidase protein. rRT-PCR was then performed and evaluated for the sensitivity, specificity and reproducibility using virus with E119V mutation and clinical samples.
RESULTSThis study described the validation of a highly sensitive and specific duplex rRT-PCR for detection of substitutions leading to the E119V amino acid change in NA protein of influenza A(H3N2). Fluorescence signals could be detected even when diluted a A (H3N2) virus (HA = 8) into 10(-5) and linear correlation between the logarithm of the viral titer with the Ct values was observed. In addition, the assay was highly specific in that there was no cross-react with other respiratory viruses, nor did two TaqMan-MGB probes. E119V substitution in quasispecies with both sensitive and resistant viruses could be detected as well. The limit of detection was 5% for quasispecies with high concentrations and 50% for quasispecies with low concentrations. The average coefficient of variation (CV) for within-run assays was 2.32% and 0.57% for H3N2-119E and H3N2-119V primer/probe sets separately, 1.77% and 0.97% for average CV of between-run assays, which exhibited good repeatability. Sequence analysis of twenty NA genes verified glutamic acid (E) at amino acid site 119, which was in consistent with the results from our rRT-PCR method.
CONCLUSIONThe assay developed in this study is highly sensitive and specific, and easy to operate; thereby it could be used for identification of A(H3N2) virus with E119V amino acid change in NA protein.
Amino Acid Substitution ; Drug Resistance, Viral ; Influenza A Virus, H3N2 Subtype ; drug effects ; enzymology ; genetics ; Mutation ; Neuraminidase ; genetics ; Nucleic Acid Probes ; Reverse Transcriptase Polymerase Chain Reaction ; methods
            
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