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Objective To discuss the change trends of pathogen of severe hand,foot and mouth dis-ease(HFMD) in Chaoshan area during 2011 to 2015. Methods All 1410 throat swabs of cases who were diagnosed as HFMD were collected from children hospitalized in our hospital during May 2011 to August 2015. Enterovirus were detected by nest RT-PCR,and the results of these positive cases diagnosed as severe HFMD were analyzed. Results (1) There were 216 positive cases(67. 29％,216/321) diagnosed as severe HFMD,including 53. 70％ ( 116/216 ) enterovirus 71 ( EV71 ), 19. 91％ ( 43/216 ) coxsackievirus A16 (CA16),12. 04％(26/216) CA6,8. 80％(19/216) CA10,3. 24％(7/216) CA4,0. 93％(2/216) coxsack-ievirus B5, 0. 46％ ( 1/216 ) enteric cytopathogenic human orphan virus and 0. 93％ ( 2/216 ) unclassified samples were unclassified to species. (2) Five cases of critical HFMD were all caused by EV71. (3) The EV71 positive samples were given priority to severe cases ( 51. 79％,116/224 ) and the non EV71 positive samples were given priority to mild cases ( 82. 08％, 458/558 ) , the difference was statistically significant (χ2 =91. 68,P<0. 001). (4) The change trends of severe HFMD year by year were consistent with the change trends of EV71 composition,and were highly correlated(Rs=0. 9,P=0. 037). (5) Severe HFMD caused by non EV71 virus gradually increased. Conclusion Severe HFMD in Chaoshan area during 2011 to 2015 were mainly caused by EV71,non EV71 viruses including CA16,CA6,CA10,CA4,coxsackievirus B5, enteric cytopathogenic human orphan virus 6 could also develop to severe HFMD. The composition ratio of severe HFMD increased accordingly in the year of EV71 as the dominant pathogen. The proportion of severe HFMD caused by non EV71 virus gradually increased after 2013 year.
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Objective To investigate the best experimental scheme of Mycoplasma solid culture combined with liquid culture.Methods A total of 961 samples of urogenital tract excretion were collected from March 2016 to June 2016.Both solid culture and liquid culture were performed for detection of Mycoplasma,false positive broths were picked out after 48 h,20 μl of each one was transformed to solid media for subculture,final results were read after 48 h.The experimental data was analyzed to find an optimal combination culture scheme.Results The positive rate of solid culture was 38.7％ (372/961) for UU and 7.8％ (75/961) for MH.The positive rate of liquid culture was 45.27％ (435/961) for UU and 7.08％ (68/961) for MH.The different rate between both methods was 9.47％ (91/961) for UU (x2 =43.61,P＜0.005),and 3.02％ (29/ 961) for MH (x2 =1.24,P＞ 0.05).The different rate was 53.5 ％ (46/86) between primary solid culture and subculture.The positive rate of total (primary solid culture and subculture) solid culture was 41.9％ (403/961) for UU and 8.6％ (83/961) for M H,which produced higher sensitivity than single (primary solid culture or subculture) solid culture.The different rate between the total solid culture and the liquid method was 6.24 ％ (60/961) for UU (x2 =17.067,P＜0.05),and 3.43 ％ (33/961) for MH (x2=5.94,P＜0.05).Conclusion Perform both solid culture and liquid culture for Mycoplasma,pick out those false positive broths for subculture.Total solid culture could be more reliable as final result,for which could combine specificity of solid culture with sensitivity of liquid culture.
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Objective To discuss the significance of different types of human rhinovirus (HRV) as pathogen and the clinical features of different types of HRV in pediatric intensive care unit(PICU).Methods Eight hundred and fifty-two nasopharyngeal aspirates specimen (NPA) were collected from children who were admitted to PICU,the Second Affiliated Hospital of Shantou University Medical College from November 2010 to October 2015 and were tested by using nested reverse transcription-polymerase chain reaction (RT-PCR).Gene fragments for VP4/VP2 capsid protein amplified from HRV positive specimens were sequenced for HRV genotype confirmation.Then clinical characteristics of these HRV positive cases were analyzed.Results Among these 852 specimens tested,214 (25.12％) were HRV positive,including 95 samples(44.39％) positive for HRV-A,17 samples (7.94％) for HRV-B,and 55 samples(25.70％)for HRV-C determined by sequence analysis;while the species of 47 samples (21.96％) of the total were unclassified clearly.HRV-A,HRV-B,HRV-C co-infection with other respiratory viruses accounted for 33.68％ (32/95 cases),29.41％ (5/17 cases),and 29.09％ (16/55 cases),respectively.The clinical characteristics of children infected with HRV-A,HRV-B,HRV-C were similar,and wheezing and polypnea were more common with HRV-C infections than HRV-A and HRV-B infections.The severity among children positive for different groups HRV showed no significant difference (H =0.631,P ＞ 0.05),as well as that between children co-infected with HRV and other viruses and those infected with HRV only (H =0.886,P ＞ 0.05).Conclusions Different types of HRV were major causes of infectious disease in pediatric critical disease.The clinical characteristics of children infected with HRV-A,HRV-B,HRV-C were similar.Wheezing and polypnea were more common with HRV-C infections than HRV-A and HRV-B infections.
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Objective To explore the morbidity and clinical characteristics of mumps meningoencephalitis in children without parotitis.Methods Two hundred and twenty-three cerebrospinal fluid (CSF) specimens were collected from children who were diagnosed as viral encephalitis at Department of Pediatrics,the Second Affiliated Hospital of Shantou University Medical College from June 2010 to February 2016.Multiplex PCR was applied to detect the mumps virus,and other common viral,including measles virus,enterovirus,enterovirus 71 type,coxsackie virus A16 type,dengue virus,Japanese encephalitis virus,rubella virus,herpes simplex virus,human cytomegalovirus,Epstein-Barr virus,Chikungunya virus and Charon evagatus in mumps virus positive specimens were detected by PCR.The clinical data of patients with mumps virus infection were analyzed.Results In 223 CSF specimens,positive mumps virus were detected in 11 cases (4.9％),of whom,the mycobacterial,fungal,conventional CSF cultures and other common viral cause in CSF were negative.One case presented parotitis on the sixth day after admission.Of 11 cases with positive mumps virus,there were 10 cases without parotitis.The cardinal symptoms of mumps meningoencephalitis in children without parotitis were fever,headache,vomiting and seizures,and the CSF parameters,brain magnetic resonance imaging,electroencephalogram ofthe patients were all similar to other viral encephalitis,while the prognosis was good in children with mumps meningoencephalitis without parotitis,but the CSF return to normal needed a long time,the longest time up to 4 weeks.Conclusion Mumps meningoencephalitis may occur in children without parotitis,and the most common symptom are fever,headache,vomiting and seizures.
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OBJECTIVE@#To assess the value of endobronchial ultrasound elastography in the diagnosis of mediastinal and hilar lymph node metastasis in lung cancer. @*METHODS@#A total of 40 patients with lung cancer underwent ultrasonic bronchoscope examination before operation. Elastography and standard endobronchial ultrasound (EBUS) of lymph nodes were performed before EBUS-guided transbronchial needle aspiration (EBUS-TBNA). The elastography characteristics was compared between benign and malignant lymph nodes. The diagnosis accuracy in malignant lymph nodes was also compared between the elastography and the standard EBUS. The value of the elastography was assessed in distinguishing the benign and malignant lymph nodes. @*RESULTS@#1) The significant indicators of standard EBUS in diagnosis of malignant lymph nodes were hypoechonic nodes, uneven echo, distinct boundary and short diameter greater than 1 cm (all P<0.01). 2) There was significant difference in the elastosonography grading score between benign and malignant lymph nodes (P<0.01). 3) The elastography grading score was more sensitive and specific in determining the malignant lymph node than the standard EBUS criteria. The area under the receiver operating characteristic curve (AUC) was maximal when the elastography grading score was ≥2.5. The specificity, sensitivity, positive predictive value, negative predictive value of elastography grading score was 76.9%, 85.7%, 85.7% and 76.9% in distinguishing malignant and benign nodes. The overall accuracy of elastography grading score was 82.3%. The combination of elastography grading score, low echo, distinct boundary and short diameter greater than 1 cm showed the best diagnostic efficiency value. The AUC was 0.911. In distinguishing malignant and benign nodes, the specificity, sensitivity, positive predictive value, negative predictive value and accuracy of the combined indexes was 84.6%, 88.1%, 90.2%, and 81.5% respectively. The overall accuracy was 86.8%. @*CONCLUSION@#The endobronchial ultrasound elastography can effectively distinguish the mediastinal and hilar lymph node metastasis in lung cancer. The diagnosis accuracy of elastography in malignant lymph node is higher than that of standard EBUS criteria. The combination of elastosonography grading score and standard EBUS criteria can improve the diagnostic efficiency.
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Humans , Bronchoscopes , Elasticity Imaging Techniques , Lung Neoplasms , Pathology , Lymph Nodes , Pathology , Lymphatic Metastasis , Diagnostic Imaging , Mediastinum , Pathology , Retrospective Studies , Sensitivity and SpecificityABSTRACT
Objective By prokaryotic expression and purifying the human bocavirus recombinant protein VP2, to establish the indirect enzyme-linked immunoassay for detection of virus. Methods We amplified the human bocavirus recombinant protein VP2 gene fragments from WHL-1 template by PCR , and cloned into the expression vector pET28a, then conversed into the BL21 (DE3) and expressed the fusion protein detected by Western Blot detection , the obtained the antibody and detected the human bocavirus in serum in Guanghzhou area in healthy people. Results The Recombinant prokaryotic expression identified correct by double enzyme, and it could occur specific reaction with the virus positive serum. The best optimal antigen coating concentration were serum multiples and blocking BSA was 2 mg/mL , 1 ∶ 200 and 1%. The best working dilution of enzyme-labeled secondary antibody was 1 ∶ 4 000. The best working hours was 1h. This detection method had good specificity and reproducibility. The cut-off of the indirect ELISA method was 0.1 and the sensitivity and specificity of the developed ELISA method were 92% and 98% respectively. The coincidence rate of determination results by the developed kit and control kit was 97%. Conclusion The competitive ELISA established by prokaryotic expressing and purifying the human bocavirus protein VP2 protein , provides a basis in detecting the human bocavirus serum antibody.
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TOF-MS has been more and more widely used in clinical and scientific research due to its advantages such as fast and easily operated.Diabetic nephropathy is the most common complications of diabetes.But clinical existing detection meth-ods to some extent have some hysteresis.The applications and developments of TOF-MS coupled with other proteomics technologies provided new ideas and methods for early diagnosis and prognosis of diabetic nephropathy.In this paper,the lat-est research results in nearest years about the application of TOF-MS to diabetic nephropathy will be reviewed.
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Objective To discuss the clinical value of the expression level of acute lower respiratory tract Boka virus (HBoV)in‐fectecd children whose detection serum specific antibody .Methods 904 cases of children with acute lower respiratory tract Boka vi‐rus infection hospitalized from March 2011 to July 2014 who were selected as study objects ,serum ,sputum ,bronchoalveolar lavage fluid HBoV DNA positive were as the gold standard for diagnosis of acute lower respiratory tract infection in HBoV ,the positive serum HBoV antibody of HBoV in children with acute lower respiratory tract infection was defined as the observation group ,serum HBoV antibody negative acute lower respiratory tract infection in children with HBoV was defined as the control group ,the correla‐tion between serum HBoV antibody and acute lower respiratory tract HBoV infection children whose clinical characteristics were analyzed .Results Serum HBoV antibody in the diagnosis of acute lower respiratory tract infection of HBoV whose sensitivity ,spe‐cificity ,positive predictive value ,and negative predictive value ,accuracy of diagnosis were separately 60 .32% 、90 .25% 、31 .67% 、96 .81% 、88 .16% .In the general data ,between the observation group and the control group in gender ,age ,hospitalization time , there were no significant differences(P>0 .05) .In the clinical manifestations ,nasal congestion and runny nose ,cough ,fever ,vomi‐ting and diarrhea ,shortness of breath ,breathing difficulties whose occur rates had no significant differences between the observation group and the control group(P>0 .05) ,the incidence of wheezing of the observation group was significantly higher than that of the control group ,the difference had statistical significance (P0 .05) .Conclusion Serum HBoV antibody is in favor of acute lower respiratory tract infection of HBoV in the diagnosis of exclusion ,and the serum HBoV antibody positive and acute lower respiratory tract infection of HBoV have a certain relationship in children with wheezing symptoms .
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Objective To discuss the correlation between the level of serum cystatin C and lipids in patients with system lupus erythematosus.Methods Used automatic biochemical analyzer to detect serum cystatin C (CysC),triglyceride (TG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C)and hsCRP levels in 136 cases of SLE patients and 113 cases of healthy people.Data obtained using SPSS13.0 software to carry on sta-tistical analysis.Results Outcome of SLE patients group compared with healthy controls,hsCRP (13.5 ± 4.85 mg/L vs 2.03±0.88 mg/L),CysC (2.63±1.95 mg/L vs 0.85±0.37 mg/L),LDL-C (3.06±1.21 mmol/L vs 2.33±0.41 mmol/L),TC (5.32±2.63 mmol/L vs 4.02±1.67 mmol/L)and TG (1.92±0.83 mmol/L vs 1.44±0.8 mmol/L)were signifi-cantly higher the difference between groups was statistically significant(t=2.45~12.4,P <0.05).Compared with healthy controls,HDL-C (1.12±0.31 mmol/L vs 1.52±0.85 mmol/L)was decreased (P <0.01).In SLE patients group,the ser-um CysC level and hsCRP,TC,TG and LDL-C were positively correlated,and the level of HDL-C was negative to the level of CysC.The health control group was no significant correlation.Conclusion Serum lipid levels of SLE patients were posi-tive to the level of CysC.Suggest that joint detection of SLE patients serum CysC and blood lipids index is helpful to the di-agnosis of SLE treatment and condition monitoring.
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Objective To investigate the relationship between Hepatitis B patients with different viral loads and immunoglob-ulin A,G,M and complement C3,C4.Methods Firstly,followed by real-time fluorescence quantitative PCR detection 210 cases of hepatitis B patients with HBV-DNA levels,according to 10n copies/ml different viral load detection results,it was divided into 102 ~108 copies/ml of the experimental groups.Then the experimental groups and control group were simulta-neously detected in immunoglobulin A,G,M and complement C3,C4.Analysed the correlation between HBV loads and im-munoglobulin A,G,M and complement C3,C4.Results When the viral loads of hepatitis B patients were 105 ~108 copies/ml,the testing results of IgA,IgG and IgM were both increasing (U =12.43,10.96,6.42,P <0.01),while C3 and C4 were both decreasing (U =8.37,6.0,P <0.01).When the viral loads of hepatitis B patients were 102 ~ 104 copies/ml,only IgA and IgM were increasing (U =2.36,2.04,P <0.05),the other testing results had no statistical significance.Between the test of 7 experimental groups compared with each other,only 104 group and 105 group had significantly changed (IgA and IgM were increasing,C4 was decreasing,U =2.39,2.46,2.09,P <0.05,IgG was increasing,U = 3.25,P <0.01),but between other low viral loads or high viral loads were not significantly differences.Conclusion The different viral loads of hepatitis B patients could cause the different changes of immunoglobulin A,G,M and complement C3,C4,especially in the 4 groups from 105 to 108 copies/ml.Followed by increasing in viral loads,there were immunoglobulin A,G,M increasing and comple-ment C3,C4 decreasing,and also serious impaction on the immune function of organism.There was a positive correlation be-tween viral loads in vivo and immune damages,correlation coefficient (γ =0.967,P <0.01).When the viral loads from 104 to 105 copies/ml,the testing results had changed significantly.It suggest that should control viral loads under 104 copies/ml in the hepatitis B antiviral treatment,so the effect of immune function damage will be the minimum.
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Objective To discuss the etiology and clinical characteristics of human rhinovirus (HRV) as pathogen of central nervous system infection .Methods Two hundred and five cerebrospinal fluid (CSF) specimens were collected from children with fever and convulsions who were admitted to the pediatric intensive care unit with suspicion of central nervous system infection from June 2011 to December 2012 .Genome Lab Genetic Analysis System (GeXP) was applied to detect HRV from CSF .Specimens with positive results were amplified by nested reverse transcription‐polymerase chain reaction and followed by gene sequencing . Clinical data of HRV positive cases were analyzed . Results Of the 205 CSF specimens ,7 samples were positive for HRV ,which were composed of 2 HRV‐A ,1 HRV‐B and 4 HRV‐C (including 1 HRV‐Ca) .There were 6 boys and 1 girl among the 7 positive cases for HRV .Six children were less than 3 years old ,except one was 9 years old .The onset time was mainly concentrated between September and October . The main clinical manifestations were fever and convulsions . The clinical diagnosis before the pathogen confirmation included viral encephalitis ,epilepsy ,febrile convulsion ,benign infantile convulsions with mild gastroenteritis (CwG ) and hand‐foot‐and‐mouth disease ( HFMD ) . Although the disease severity of the 7 cases varied ,all ended with favorable prognosis .Conclusions HRV is one of pathogens of viral central nervous system infection .All types of HRV can cause central nervous system infection ,among which HRV‐C accounts for the majority .The clinical manifestations of HRV central nervous system infection could mimic febrile convulsion ,CwG and HFMD .
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Objective To understand the pathogen of viral encephalitis(VE)in children,and establish a rapid and specific method for detecting human rhinovirus(HRV),and investigate the correlation between HRV and viral encephalitis(VE). Methods 169 CSF specimens were collected from children with convulsions and fever,who were admitted to the pediatric intensive care unit (PICU)of Second Affiliated Hospital of Shantou University Medical College between January 2012 and December 2012.Nested RT-PCR was used to detected HRV in CSF specimens,and the positive PCR products were se-quenced,then analyzed and constructed the phylogenetic tree by software.Results 39 (23.1%)out of 169 samples were HRV positive.Among them,148(87.6%)children were below 5 years old.The detection rate of HRV increases from July to September,and reached its highest point in September.Sequence analyzed showed that the 39 HRV positive specimens inclu-ding 18(46.1%,18/39)positive for HRV-A,7(17.9%,7/39)positive for HRV-B,14(35.9%,14/39)positive for HRV-C. There were 8 out of 28 VE cases were detected in HRV,including 3(50%,3/6)positive for HRV-C.Conclusion HRV could be detected in CSF specimens by nested RT-PCR,including three types of HRV,combined with clinical symptoms con-sidered that HRV may be one of the VE pathogen.
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Objective To study the significance of human rhinovirus C as a pathogen and the clini-cal features of human rhinovirus C infection in pediatric intensive care unit. Methods From November 2010 to April 2012,570 nasopharyngeal aspirates specimens were collected from children who were admitted to the pediatric intensive care unit with respiratory infections. Nest reverse transcription-polymerase chain reactions were applied to detect the human rhinovirus C. The other common respiratory viruses were detected by multi-plex polymerase chain reaction. The clinical data were collected. Results One hundred and seventy human rhinovirus positive samples ( 29. 8%) were detected in 570 nasopharyngeal aspirates specimens. The VP2/VP4 and 5′UTR region of the human rhinovirus genome was amplified from 170 human rhinovirus positive samples with 80. 6%(136/170) success. While 20. 0%(34/170) samples in total were unclassified to spe-cies. There were 85 single infected samples including 52 of type A,7 of type B,26 of type C. The nucleotide homology was 74. 0% to 99. 2% and the nucleotide variations was 3. 4% to 32. 3% in stains of human rhino-virus C. The late fall and early winter were the epidemic seasons of human rhinovirus C infection. Cough,fe-ver, polypnea and wheezing were the common symptoms. Conclusion Human rhinovirus C is the major cause of infectious disease in pediatric critical illnesses. Human rhinovirus C infections often cause cough, fever,polypnea and wheezing.
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Objective To analyze the status of human metapneumovirus in children with respiratory tract infection in Shenz-hen and Shantou and the clinical characteristics of the diseases with hMPV infection.Methods The viral nucleic acid of 1 217 samples were amplified by RT-PCR(1 137 from children with respiratory tract infection,80 from health children),and five positive PCR products were chosen randomly for sequencing.Nucleotide sequences alignment and phylogenetic analysis were performed with DNAstar software.The clinical data of the children with hMPV infection were analyzed.Results The positive rate of hMPV infection identified from 1 137 children wtith respiratory tract infection was 4.49% (51/1 137), hMPV was significantly prevalent in February,March and April.Most of the positive cases was under 3 years old.Five hMPV gene fragments of sequence were closely related to hMPV in the GenBank.Similarity of hMPV partial N gene with which published at nucleotide levels varied from 80.8%~98.4%.Phylogenetic tree of nucleotide revealed two different gen-otypes.All hMPV positive patients suffered from fever,cough and wheezing.The clinical diagnoses were wheezing pneumo-nia (17),bronchiolitis (9).None of the nasal swabs from 80 healty subjects were tested hMPV specific gene fragment.Con-clusion hMPV is an important viral pathogen in children with respiratory tract infection in Shenzhen and Shantou.Two genotypes may circurate in these areas.
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Objective To evaluate the value of HbA2 level determined by capillary electrophoresis (Hb-CE)in screening and diagnosis of thalassemia.Methods HbA2 level of 249 thalassaemia carriers and 142 healthy controls confirmed by molecular biological detection were determined by Hb-CE method.The thalassaemia carrier subjects were divided into different groups and subgroups according to their results of gene detection.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value for the diagnosis ofα-thalassemia,β-thalassaemia,α,β-thalassaemia were calculated under different HbA2 cut-off value.Results Mean value of HbA2 in healthy controls was (3.03±0.27)%.Mean values of HbA2 inα-thalassemia group and its subgroups of silentα-thalassemia,standardα-thalassemia and hemoglobin H disease were (2.38± 0.55)%,(2.61±0.46)%,(2.47 ± 0.32)% and (1.07 ± 0.17)%,respectively.Mean values of HbA2 inβ-thalassaemia group and itsβ0 subgroup,β+ subgroup were (5.65±0.47)%,(5.71±0.48)% and (5.56±0.43)%.Mean value of HbA2 in compoundαandβ-thalassaemia group was (5.7±0.82)%.Compared with healthy controls,HbA2 level inα-thalassemia group,silentα-thalassemia subgroup,standardα-thalassemia subgroup and hemoglobin H disease group decreased signifi-cantly (t values of 11.73,5.02,12.91 and 33.46,respectively,P0.05).HbA2 level inβ-thalassaemia group,β0 subgroup,β+ subgroup and compoundαandβ-thalassaemia group increased significantly (t values of 55.12,44.33,38.94 and 9.10,respectively,P0.05).Of 249 thalassemia carriers,all 124β-thalassaemia carriers were distinguished with ele-vated HbA2 level (>3.5%)determined by Hb-CE and only 57 were distinguished from 117α-thalassemia carriers by Hb-CE.Under the cut-off value of 2.5%,the sensitivity,specificity,positive predictive value,negative predictive value and accu-racy for the diagnosis ofα-thalassemia were 48.72%,97.18%,93.44%,69.70%,75.29%,respectively.Under the cut-off value of 3.5%,they were 100.00%,98.59%,98.41%,100%,and 99.25% for the diagnosis ofβ-thalassaemia,respectively. The analysis of ROC curve showed that the optimal HbA2 cut-off values for diagnosis ofα,β-thalassaemia by capillary elec-trophoresis were 2.8% and 3.7%,respectively.Conclusion When no abnormal bands,the elevated HbA2 (>3.7% in this study)determined by Hb-CE could be used as a marker forβ-thalassaemia diagnosis,but theβ-thalassaemia co-existingα-thalassemia could not be differentiated fromβ-thalassaemia diagnosis.Decreased HbA2 level (<1.5% in this study)and HbH band could be used for the diagnosis of hemoglobin H disease.Only HbA2 determination by Hb-CE has no clinical sig-nificance for the screen and diagnosis ofα-thalassemia.
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Objective To analyze the clinical features of PICU patients with bocaviral infection.Meth-ods Nasopharyngeai aspirates specimens were collected from 450 children who were admitted to PICU with a-cute respiratory tract infection in our hospital from June 2010 to December 2011 .Multiplex PCR was applied to detected human bocavirus and emerging respiratory virus.Bocavirus positive PCR results were sequenced and the clinical data of the positive cases were analysed.Results Human bocavirus positive samples were detected in 30 cases(6.7%) among 450 throat swab specimens.Human bocavirus as a single infection was found in 16 cases (53.3%).Mixed infections were found with in 14 cases(46.7%) of 30 positive samples.According to pediat-ric critical illness score,there were 13 cases of non-serious,2 cases of serious and 1 case was extremely serious in 16 single infections cases.There were 12 cases of non-seriuo s and 2 serious cases in 14 mixed infections. There were no statistically significant differences between single and mixed infections in the severity of the dis-ease( P>0.05 ) .Conclusion Bocavirus can cause severe respiratory tract infections.Mixed infections does not increase the severity of the disease.
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The severe infectious diseases caused by virus are occurring with increasing frequency, which poses a rising global threat to human health and life.There are many kinds of viruses that cause a wide variety of viral diseases.The same kind of virus is able to cause different diseases, meanwhile a disease can be caused by different viruses.All these conditions make the relationship between viral pathogens and infection diseases complicated.At present, no effective laboratory detection methods for most of virus infectious diseases are developed.But the rapid development of molecular diagnostic technologies makes it possible for clinical laboratory to detect viral infection diseases rapidly and simultaneously.This review summarized the present and development perspectives of laboratory tests for the diagnosis of clinical virus infections, attempting to give some advices for laboratory diagnosis procedure of viral infections.
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Objective To express the capsid proteins of WU polyomavirus(WUPyV) for research and find antigen for diagnostic value. Methods Coding sequences of capsid proteins of WU polyomavirus by PCR were cloned in prokaryotic expression vector PGEX-20T. Recombinant plasmids were transformed into E. coli BL21(DE3) and induced by IPTG for proteins expression. Recombinant proteins were identified by Western blot. Results SDS-PAGE proved that recombinant proteins showed three bands with molecular relative mass of 69×103, 63×103 and 56×103. The recombinant proteins were recognized by anti-GST McAb. The antigenicity was tested by Western blot using 16 WU polyomavirus positive and 70 negative sera. Conclusion Recombinant VP1, VP2 and VP3 expressed in E. coli can combine with WUPyV-Ab and have good antigenicity. They can be used for further research.
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Objective To evaluate five detection methods for the laboratory diagnosis of Clostridium difficile infection in the hospitals of USA, and explore a sensitive, specific, accuracy and rapid regimen for the early diagnosis of Clostridium difficile infection. Methods A total of 174 stool specimens submitted to the clinical microbiology laboratory for Clostridium difficile testing were separately tested by five methods including toxigenic culture (TGC), Premier Toxin A&B EIA( A/B-EIA), C. Diff Quick Chek Complete( DEIA), BD G eneOhm Cdiff assay(BD-PCR) and Laboratory-developed PCR(LD-PCR). The gold standard of TGC was used as a reference criterion, and the sensitivity, specificity, positive predictive value ( PPV )and negative predictive value (NPV) of A/B-EIA, D-EIA, BD-PCR and LD-PCR assays were determined. Results Among the 174 specimens studied, 24 were defined as true positives for Clostridium difficile infection by TGC assay, giving a positive rate of 13.8% (24/174). In comparison to the standard,the sensitivity, specificity, PPV and NPV were 62.5%, 99.3%, 93.8% and 94.3% for A/B-EIA;66.7%, 98.7%, 88.9% and 94.9% for D-EIA; 83.3%, 98.7%, 90.9% and 97.4% for BD-PCR;79.2%, 93.3%, 65.5% and 96.6% for LD-PCR. Among all tested specimens, 34 were positive by atleast one of five methods, and of which 15 were concordant by all five methods. The D-EIA results were divided into three groups depending on results of GDH and (or) toxins A/B: 18 were positive for both GDH and toxins A/B, 23 were positive for only GDH, and 133 were negative for both GDH and toxins A/B. Of 18 positive specimens by D-EIA assay, all were concordant with results of BD-PCR assay and 16 were agreement with results of TGC assay. Twenty-two of 24 positive specimens by TGC assay were included in 41 specimens that were positive for GDH. Among eight false negative specimens by D-EIA assay, four were differentiated as positive results by BD-PCR. According to the present study, the sensitivity, specificity,PPV and NPV of a two-step detection algorithm in combination with D-EIA and BD-PCR assays were 83.3%, 98.7%, 90.9% and 97.4%, respectively. Conclusions From the point of technological evaluation, BD-PCR is preferable. A two-step detection algorithm combining D-EIA with BD-PCR is proposed for the laboratory diagnosis of Clostridium difficile infection. This algorithm has demonstrated an excellent sensitivity and specificity, as well as decreased test turnaround time and test cost.
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Objective To investigate the prevalence of human bocavirus(HBoV)among children with acute respiratory tract infection(ARTI)in Guangdong Province.Methods Four hundred and forty-seven nasopharyngeal aspirates or swabs samples from children with ARTI in Guangdong Province were collected from June 2007 to May 2008.HBoV capsid protein VP gene fragments were detected using polymerase chain reaction(PCR).Positive PCR products were sequenced.The DNA and translated amino acid sequences were aligned with known HBoV sequences in GenBank and phylogenetic analysis was also done.Results The positive rate of HBOV was 5.1%of samples from 447 ARTI cases.Ten samples were positive for both HBoV and other respiratory virus,which was 43.5%of positive samples.The main diagnosis for HBoV positive children included wheezing pneumonia,bronchiolitis and bronchial pneumonia.HBoV positive children ranged from 42 days to 6 years old,and most of them were younger than one year.HBOV infection was more common during summer,early autumn and late spring.Through sequence alignment and phylogenetie analysis,the DNA sequences and amino acid sequences of VP gene fragments of isolated HBoV strains showed 97.8%-98.8%and 99.3%-100.0%identity with ST1,respectively.Conclusions HBoV is one of the important pathogens of lower respiratory tract infection in children in Guangdong Province,which is more prevalent in infants younger than one year.Although VP gene fragment of HBoV is conservative in general,there are still some missense mutations.