Identification of Respiratory Syncytial Virus A and B Subtypesby Tube Double Nested Polymerase Chain Reaction and Gene Sequencing Technology / 现代检验医学杂志

Objective Objective to establish a method for identification of respiratory syncytial virus (RSV) A and B subtypes for clinical research and application.Methods Using fluorescent quantitative polymerase chain reaction (FQ-PCR) screened 50 cases of throat swab that RSV were positive in hospitalized children from 2015 to 2017.The genotyping was performed according to the nucleotide sequence of G protein coding gene,and a single tube double nested PCR primers was designed for it.A and B subgroup by sequencing to conduct comparative analysis with nucleotide sequence in the Genebank.The results were analyzed by chi-square test.Results In the 50 cases of throat swab RSV positive children,respiratory syncytial virus A and B subtype and mixed infection rates were 82.00％,14.00％ and 4.00％,respectively.The difference was statistically significant (x2=81.06,P＜0.01).The RSV fractal results were consistent with the gene sequencing results.Conclusion The eastern part of shenzhen was dominated by respiratory syncytial virus A subtype epidemic and mixed infection.Single tube double nested polymerase chain reaction (PCR) and gene sequencing technique is suitable for the identification of A and B subtypes of RSV.It is characterized by high sensitivity,specificity and high accuracy.

Relationship between M-Protein of Multiple Myeloma and False Positive Syphilis Serological Results / 中国实验血液学杂志

<p><b>BACKGROUND</b>The false positive in conventional syphilis serological test was found in patients with multiple myeloma (MM).</p><p><b>OBJECTIVE</b>To investigate the relationship between the M-protein of patients with MM and the false positive in conventional syphilis serologic test.</p><p><b>METHODS</b>The M-protein of 68 MM cases was typed with immunofixation electrophoresis and 68 cases of MM were screened with non-specific and specific syphilis serologic tests, then the samples with syphilic serological positive were chosen and confirmed with immonobloting test, finally the relationship between M protein of MM and the false positive of syphilis serological test were analysed.</p><p><b>RESULTS</b>Four out of 68 cases showed the positive in syphilis serological test and further were confimed to be false positive by immunoblotting test, the false positive rate was nearly 6%. The M-protein of MM patients in our hospital mostly possessed IgG, κ type, followed by IgA, κ type, light chain κ type. In general, κ : λ = 2.4 : 1. Among samples of 4 cases with syphilis serological positive 2 cases were of IgG and κ type, 1 case was of IgG, λ type, another 1 case was IgA, κ type.</p><p><b>CONCLUSION</b>The M-protein of IgG and IgA types in MM patients results in syphilis serological false positive reaction. The clinicians and laboratorial technicians should pay a great attention to screen the MM patients for the false positive syphilis serological test so as to avoid the misdiagnosis and subsequent embarassment.</p>

Monitoring of viral pathogens in pediatric intensive care unit and analysis of clinical significance / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the characteristics of viral spectrum and clinical features of children in pediatric intensive care unit (PICU).</p><p><b>METHOD</b>Nasopharyngeal aspirate specimens (NPA) from 349 patients(1 from each) and 130 cerebrospinal fluids (CSF) specimens were collected from children who were admitted to the PICU of Second Affiliated Hospital of Shantou University Medical College. Additional 87 NPA specimens were collected from healthy children for routine examination on the physical examination center, and the clinical data were collected. Multiplex PCR was applied to detect 16 kinds of viruses from NPA and CSF. Fluorescence quantitative PCR was applied to detect 13 viruses from CSF and to analyze the clinical data of positive cases.</p><p><b>RESULT</b>There were 209 samples (59.9%) of the 349 NPA specimens were positive for viruses, which included 117 cases positive for human rhinovirus (HRV), 60 for respiratory syncytial virus (RSV), 20 for influenza virus A (Inf A), 10 for adenovirus (ADV), 6 for parainfluenza virus type 3(PIV-3), 6 for human Boca virus (HBoV), 5 for influenza virus C(Inf C), 4 for parainfluenza virus type 4(PIV-4), 4 for human coronavirus-HKU1/OC43, 3 for influenza virus B (Inf B), 3 for WU Polyomavirus (WUPyV), 2 parainfluenza virus type 1(PIV-1), 2 human metapneumovirus (HMPV) and 1 human coronavirus-NL63/229E. But none from 87 healthy controls were positive for any respiratory virus. Among the 130 CSF specimens, in 58 cases the diagnosis was viral encephalitis. There were 22 samples (37.9%) among the 58 CSF specimens positive for viruses, which included 14 enterovirus (EV), 3 human cytomegalovirus (HCMV), 2 mumps virus, 1 coxsackie virus A16 (Cox-A16), 1 herpes simplex virus (HSV) and 1 human rhinovirus (HRV). The total positive rate was 63.3% (221/349) . Co-infection by at least 2 viral pathogens under study was observed in 45 of the 349 patients (12.9% of the total number of cases, 20.4% of the positives cases). The commonest pathogens in co-infected samples were WUPyV (100%) and HMPV(100%). The positive rate of virus peaked in the first 6 months of life, the rate in boys were higher than in girls and the peak season was summer. The numbers of none serious cases in the virus positive group were less than those in the virus negative group while the numbers of extremely serious cases in the virus positive group were higher than in the virus negative group.</p><p><b>CONCLUSION</b>Viral pathogen is a major cause of infectious disease in pediatric critical illnesses and virus infection may lead to severe illness.</p>

Detection and clinical features of human rhinovirus in hospitalized children with acute respiratory tract infection in eastern areas of Guangdong province / 中华流行病学杂志

Objective To investigate the prevalence and clinical features of human rhinovirus (HRV) infection in hospitalized children with acute respiratory (ARI) in eastern areas of Guangdong province from 2008 to 2010.Methods From Oct.2008 through Sep.2010,nasopharyngeal aspirates were collected prospectively,from hospitalized children with acute lower respiratory tract infection at the Second Hospital,affiliated to the Shantou University Medical College.Multiplex PCR was applied to detect ten kinds of viruses including HRV,RSV in the hospitalized children with respiratory tract infection.Clinical data on HRV-positive cases or RSV-positive cases were collected and analyzed.Results Among all the 1335 specimens,124 were confirmed as HRV-positive cases (9.3％),with IVA-positive rate as the highest (25.1％),followed by RSV-positive rate (15.1％).HRV infection occurred sporadically around the year,with the highest HRV-positive rate seen in spring 2009 and autumn in 2010.Symptoms,signs,chest X-ray,leukocyte count and CRP count did not differ between patients with co-infection or single HRV infection.Clinical symptoms or signs were similar between those with single HRV infection or single RSV infection in children,but the single RSV infected children were more frequently seen with wheeze and cough.28.4％ of the single RSV infected children had bronchiolitis while 10.7％ of single HRV infected children were seen (x2=0.281,P=0.596).Conclusion HRV was a relatively common cause for acute respiratory infections in the eastern areas of Guangdong province.The highest HRV-positive rate was slightly different in different years.Infants and young children were generally susceptible to rhinovirus infection.Bronchiolitis,wheeze and cough associated with HRV infection happened less than those caused by RSV.

Detecting the occult HBV infection: a laboratory study / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To see the HBV DNA detection instance in the HBsAg negative people and to study the serological method detection strategy for detecting hepatitis B virus large surface protein (HBLP) to filtrate the occult HBV infection.</p><p><b>METHODS</b>The HBsAg negative serum samples were divided into HBsAb negative and positive two species according to the hepatitis B virus markers (HBVM) in daily work excepting the special HBVM modes. Total 2000 stochastic serum samples with 1000 HBsAb negative results and 1000 HBsAb positive results were collected from the copy tubes to detect HBVM with national ELISA reagent kits and put them -20 degrees C frostily. Mixed samples (8 x 30 microl) were analyzed with fluorescence quantitative PCR (FQ-PCR) and filtrated the individual positive samples. The filtrated samples were doubly tested again with American MONOLISA HBsAg ULTRA reagents.</p><p><b>RESULTS</b>No HBV DNA positive results were found out from the 1000 HBsAb positive samples and 19 cases HBV DNA positive results were found out from the 1000 HBsAb negative samples. On these 19 samples, the HBsAg results from the American MONOLISA HBsAg ULTRA reagents were all positive and the HBLP results were all positive, too. The 19 HBV DNA quantitative results were divided into 2 cases more than 500 copies/ml, 3 cases between 400-500 copies/ ml, 3 cases between 300-400 copies/ml, 7 cases between 200-300 copies/ml and 4 cases between 100-200 copies/ml.</p><p><b>CONCLUSION</b>The leaked samples tested HBsAg with national reagents are mostly from the HBsAb negative people. HBLP results may be positive on these samples and detecting HBLP marker is propitious to filtrate the occult HBV infection. This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.</p>

Detection and clinical characterization of WU polyomavirus in acute respiratory tract infection in children / 中华儿科杂志

<p><b>OBJECTIVE</b>WU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, has been found to be associated with respiratory tract infections recently. But the role of the WUPyV as agents of human disease remains uncertain. We sought to describe the detection and clinical characterization of WUPyV in acute respiratory tract infection in children.</p><p><b>METHOD</b>From July 2008 through June 2009, nasopharyngeal aspirates were collected from 771 children who were hospitalized with acute respiratory tract infection in Second Affiliated Hospital of Shantou University Medical College, and from 82 asymptomatic children who visited the health checkup clinic. WUPyV was detected by using PCR technology and was identified by using DNA sequencing. All WUPyV-positive specimens were screened for 9 common viruses [influenza A and B, respiratory syncytial virus (RSV), parainfluenza virus (PIV) 1 and 3, human metapneumovirus, human bocavirus, adenovirus and rhinovirus] by using PCR or RT-PCR. The clinical data of WUPyV infection were collected and analyzed.</p><p><b>RESULT</b>In this study, fifteen of the 771 tested specimens with acute respiratory tract infection were positive for WUPyV, the positive rate was 1.95% and all of the asymptomatic children who visited the health checkup clinic were negative. Of the 15 cases who were positive for the virus, the age range was 2 to 48 (mean 18.8) months, 9 (60%) were male and 6 (40%) were female. WUPyV was the sole virus detected in 9 specimens (60%) from patients with acute respiratory tract infection. WUPyV was associated with the co-infection with another respiratory virus in 6 of 15 (40%) cases, most frequently with RSV (n = 4), followed by adenovirus (n = 1) and rhinovirus (n = 1). The most common clinical findings in the patients with WUPyV were cough, fever and wheezing. The most frequent diagnoses were pneumonia (n = 8), bronchiolitis (n = 4), upper respiratory tract infections (n = 2) and bronchitis (n = 1). A severe case was complicated with viral encephalitis.</p><p><b>CONCLUSION</b>WUPyV may be a respiratory pathogen because it was the sole virus detected in 9 specimens from patients with respiratory illness and all of the asymptomatic controls were negative. The most common clinical findings are cough and wheezing. Young children may be susceptible to infection with this virus and occasionally the infection with this virus may cause severe disease. More comprehensive and in-depth studies are required to prove the pathogenicity of these viruses.</p>

Detecting hepatitis B virus large surface protein to filtrate the occult HBV infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Detecting hepatitis B virus large surface protein (HBLP) with serological method to filtrate the occult HBV infection and study the clinical detection strategy.</p><p><b>METHODS</b>Two thousands HBsAg negative stochastic serum samples were collected from the copy tubes in daily work to detect hepatitis B Virus markers (HBVM) with national EHSA regent kits and put them -20 degrees C frostily. The 2000 samples were detected with HBLP and filtrated the positive samples. The HBsAg markers were doubly counterchecked with other two adding kinds of national ELISA regent kits (total 3 kinds) at the filtrated samples. The last samples were doubly tested again with American MONOLISA HBsAg ULTRA regents. HBV DNA levels were quantitative analyzed with fluorescence quantitative PCR (FQ-PCR) and taking the mean results.</p><p><b>RESULTS</b>Fifteen HBLP positive samples were detected out from the 2000 serum samples. We educed the conform negative results from the three kinds of national regents but conform positive results from the American MONOLISA HBsAg ULTRA regents in repeating HBsAg detection at the 15 samples. The HBV DNA FQ-PCR quantitative results were all less than 500 copies/mi and divided into two cases hetween 400-500 copies/nil, three cases 300-400 copies/ml, five cases 200-300 copies/ml, four cases 100-200 copies/ml and one case was less than l00copies/ml.</p><p><b>CONCLUSION</b>The false HBsAg negative results for serum samples are more generally through national regents than through importations and HBLP results mayhe are positive in these samples. Detecting HBLP marker is propitious to filtrate the occult HBV infection. This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.</p>

Viral pathogens of acute lower respiratory tract infection in hospitalized children from East Guangdong of China / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the viral pathogens of acute lower respiratory tract infection (ALRTI) in hospitalized children from East Guangdong Province of China and the relationship of the pathogens with age and seasons.</p><p><b>METHODS</b>The nasopharyngeal aspirates samples obtained from 345 hospitalized children with ALRTI were investigated for respiratory syncytial virus (RSV), human bocavirus (HBoV), human metapneumovirus (hMPV), influenza virus types A and B, rhinovirus, parainfluenza virus types 1 and 3 and adenovirus by PCR.</p><p><b>RESULTS</b>Viral pathogens were detected in 178 patients (51.6%). RSV was the most frequent (19.3%). Novel viruses hMPV (3.2%) and HBoV (3.2%) were found. A highest detection rate (61.9%) of virus was found between January to March. The infants aged 1 to 6 months showed a higher detection rate (71.3%) of virus than the other age groups. The detection rate of viral pathogens was 72.6% in children with bronchiolitis, followed by asthmatic bronchitis (70.0%) and bronchial pneumonia (44.6%).</p><p><b>CONCLUSIONS</b>RSV remained the leading viral pathogens in children with ALRTI in East Guangdong of China. Novel viruses HBoV and hMPV were also important pathogens. The detection rate of viral pathogens was associated with seasonal changes and age. Different respiratory infectious diseases had different viral detection rates, with highest detection rate in bronchiolitis cases.</p>

Expression,purification and identification of hepatitis E virus pb166-GST fusion protein / 中华检验医学杂志

Objective To express,purify and identify recombinant hepatitis E virus(HEV) pb166-GST fusion protein using GST gene fusion system and investigate its potential role in researching Hepatitis E diagnostic antigen field.Methods The recombinant E.coli BL21 performed by our own laboratory was used to induce the HEV pb166-GST expression with IPTG.The products were purified by BD Biosience GST purifying system.The specific expression was identified by SDS-PAGE and Western blot.The experiment conditions and results were described and analysed.Results The resolved HEV pb166-GST fusion protein on SDS-PAGE showed a major band at position of 43 kD.The expressed proteins had a single expected band after purify and the protein was recognized by anti-GST antibody on PVDF membrane.Conclusion The recombinant HEV pb166-GST fusion protein is expressed in recombinant E.coli BL21 efficiently in this way,and might be used as a candidate for diagnostic antigen of HEV.