[Microorganisms in the urethral secretions of chronic prostatitis patients: A genomic analysis].

OBJECTIVE: To investigate the differences in the microbial floras in the urethral secretions of the patients with chronic prostatitis to provide some reliable pathogenic evidence for the diagnosis and treatment of the disease. METHODS: Using high-throughput second-generation sequencing technology, we detected the microorganisms in the urethral secretions from 33 chronic prostatitis patients and 30 normal healthy males. We analyzed the significant differences in the microbial flora between the two groups via the rank-sum test and performed data processing with the bioinformatics software, P < 0.05 considered as with statistically significant difference. RESULTS: Statistically significant differences were observed in 17 kinds of bacteria from the urethral secretions between the normal healthy controls and chronic prostatitis patients. LEfSe analysis showed that the microorganisms with most significant abundance difference in the urethral secretions of the chronic prostatitis patients included micrococcaceae, coriobacteriaceae, coriobacteriales, coriobacteriia, weeksellaceae, comamonadaceae, enterobacteriaceae, enterobacteriales, xanthomonadaceae, and xanthomonadales. Principal component analysis (PCA) revealed significant difference in the microbial composition between the two groups. CONCLUSIONS: There is a certain correlation between chronic prostatitis and changes in the composition of urethral microbial floras. Chronic prostatitis may result from concerted action of multiple microbes rather than a single one.

[Clinical Significance of Gene Mutation Detection for Female Heterozygotes with Glucose-6-Phosphate Dehydrogenase Deficiency].

OBJECTIVE: To explore the clinical significance of G6PD gene mutation detection in female heterozygote with G6PD deficiency. METHODS: G6PD activity and fourteen common G6PD gene mutations in female blood samples were detected by biochemical phenotype detection and PCR-reverse dot blotting, respectively. Unidentified genotype of G6PD positive samples was further ascertained by direct DNA sequencing. The results from two methods were compared and analyzed. RESULTS: A total of 493 unrelated females were enrolled, and the G6PD activity and G6PD mutations was detected. Among them, 473 females were found to be normal in G6PD activity and 20 females with G6PD deficiency, and the detection rate by G6PD activity method was 4.06%. In all enrolled females, G6PD gene mutations, including the mutation of c.1311 C＞T, were identified in 130 females, and the detection rate was 26.3%. Detection rate of the mutations that can lead to G6PD deficiency was 8.11%. The detection rates between the two methods were significantly different (P＜0.01). The misdiagnosis rate of the G6PD activity detection reached 49. 94% for the female heterozygotes. Eight G6PD mutations and 13 mutation patterns were identified in the research, and most of mutation patterns were single nucleotide missense mutation. In addition to c.1311C＞T mutation, the most common mutations were c.1376G＞T, c.1388G＞A and c.95 A＞G. G6PD mutations were identified in 19 of 20 females with G6PD deficiency, and were also detected in 21 of 473 females with normal G6PD activity, of which the rate of heterozygous mutation was 90.88%. CONCLUSION: The phenotype detection based on G6PD enzyme activity alone is not sufficient for the diagnosis of female heterozygotes. The detection of G6PD mutations that covers the common mutations in specified region can effectively identify the female heterozygotes with normal G6PD activity.

[Relationship between M-Protein of Multiple Myeloma and False Positive Syphilis Serological Results].

BACKGROUND: The false positive in conventional syphilis serological test was found in patients with multiple myeloma (MM). OBJECTIVE: To investigate the relationship between the M-protein of patients with MM and the false positive in conventional syphilis serologic test. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

A 3-year prospective study of the epidemiology of acute respiratory viral infections in hospitalized children in Shenzhen, China.

BACKGROUND: The epidemiology of local viral etiologies is essential for the management of viral respiratory tract infections. Limited data are available in China to describe the epidemiology of viral respiratory infections, especially in small-medium cities and rural areas. OBJECTIVES: To determine the viral etiology and seasonality of acute respiratory infections in hospitalized children, a 3-year study was conducted in Shenzhen, China. METHODS: Nasopharyngeal aspirates from eligible children were collected. Influenza and other respiratory viruses were tested by molecular assays simultaneously. Data were analyzed to describe the frequency and seasonality. RESULTS: Of the 2025 children enrolled in the study, 971 (48.0%) were positive for at least one viral pathogen, in which 890 (91.7%) were <4 years of age. The three most prevalent viruses were influenza A (IAV; 35.8%), respiratory syncytial virus (RSV; 30.5%) and human rhinovirus (HRV; 21.5%). Co-infections were found in 302 cases (31.1%), and dual viral infection was dominant. RSV, HRV and IAV were the most frequent viral agents involved in co-infection. On the whole, the obvious seasonal peaks mainly from March to May were observed with peak strength varying from 1 year to another. CONCLUSIONS: This study provides a basic profile of the epidemiology of acute respiratory viral infection in hospitalized children in Shenzhen. The spectrum of viruses in the study site is similar to that in other places, but the seasonality is closely related to geographic position, different from that in big cities in northern China and neighboring Hong Kong.

Respiratory virus infections among children in South China.

Acute respiratory tract infection is an important cause of morbidity and mortality with a worldwide disease burden. This study aimed to determine the prevalence and clinical characteristics of children with viral-induced acute respiratory tract infection, in Southern China. Nasopharyngeal aspirate samples from 1,980 pediatric patients with suspected acute respiratory tract infection, and 82 samples from healthy subject controls were collected for routine examination at the Second Affiliated Hospital of Shantou University Medical College, from October 2007 to August 2011. Specimens were tested by multiplex polymerase chain reaction (mPCR). At least one or more viruses were detected from 1,087 samples (54.9%). These included laboratory confirmations for 446 respiratory syncytial virus (RSV), 386 influenza virus A (FluA), 315 human rhinovirus (HRV), 135 human bocavirus (HBoV), 119 Parainfluenza virus 3 (PIV3), 82 Parainfluenza virus 1 (PIV1), 66 adenovirus (ADV), 53 WU polyomavirus (WUPyV), 52 human metapneumovirus (hMPV), and 29 influenza virus B (FluB) samples. Samples from healthy subjects were negative for any virus. Of the patients with positive specimens, 107 (9.8%) were admitted to pediatric intensive care unit (PICU). Co-infection with at least two of the viral pathogens under study was observed in 325 of the 1,980 patients (16.4% of the total number of cases). These findings may help in the diagnosis of viral infections of the respiratory tract in children, and help to consider current and potential therapeutic approaches for the treatment of acute respiratory tract infection, and further respiratory complications.

[Monitoring of viral pathogens in pediatric intensive care unit and analysis of clinical significance].

OBJECTIVE: To study the characteristics of viral spectrum and clinical features of children in pediatric intensive care unit (PICU). METHOD: 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. RESULT: 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. CONCLUSION: Viral pathogen is a major cause of infectious disease in pediatric critical illnesses and virus infection may lead to severe illness.

[Detection and clinical features of human rhinovirus in hospitalized children with acute respiratory tract infection in eastern areas of Guangdong province].

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 (χ(2) = 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.

Oncogenic HPV infection resembling pseudocondyloma of vulvae: a descriptive study of 31 cases.

OBJECTIVE: Report a special type of human papillomavirus infection resembling pseudocondyloma of vulvae. The characteristics of the clinical manifestation and subtypes of the HPV infection were studied. METHODS: Thirty-one women with unprotected sex and genital warts resembling pseudocondyloma of vulvae were involved in this study. The clinical and histological manifestations were characterized. The genotypes of human papillomavirus were determined by a reverse hybridization of polymerase chain reaction (PCR) RESULTS: Thirteen HPV genotypes were detected from 31 cases. Of 13 genotypes, 3 were non-oncogenic and 10 were oncogenic. At least, one of oncogenic genotypes was detected from each patient. Among 31 cases, 10 were found with one genotype of the HPV infection, 14 with 2 genotypes of HPV and 7 with at least 3 genotypes. The most common genotype was genotype 16 that was detected from 15 of 31 samples (48%). CONCLUSION: There is a special type of oncogenic HPV infection clinically resembling pseudocondyloma of vulvae. Some of the patients who were infected with this type of HPV infection may have been misdiagnosed as pseudocondyloma of vulvae in the past time.

WU polyomavirus infection among children in South China.

This study aimed at investigating the prevalence and clinical characteristics of children with respiratory infection by WU polyomavirus (WUPyV) in Southern China. Nasopharyngeal aspirate samples were collected from 771 children with acute respiratory tract infection admitted to hospital and 82 samples from healthy subjects for routine examination at the outpatient service at the Second Affiliated Hospital of Shantou University, Medical College from July 2008 to June 2009. WUPyV was detected by the polymerase chain reaction (PCR) and DNA sequencing. All WUPyV-positive specimens were characterized further for nine viruses causing common respiratory infections, including influenza A and B, respiratory syncytial virus (RSV), parainfluenza virus (PIV) 1 and 3, human metapneumovirus, human bocavirus, adenovirus, and rhinovirus by PCR or real time (RT)-PCR. Fifteen out of 771 specimens from patients with acute respiratory tract infection, but none from healthy subjects, were positive for WUPyV and the positivity rate was 2%. Patients with WUPyV infection were between 2 and 48 months of age, and nine of the patients were male while six female. Four out of 15 patients were co-infected with RSV, one with adenovirus or rhinovirus, respectively. Patients with WUPyV infection displayed predominantly cough, moderate fever, and wheezing, and were diagnosed with pneumonia (n = 8), bronchiolitis (n = 4), upper respiratory tract infections (n = 2) and bronchitis (n = 1). One patient developed encephalitis. Therefore, WUPyV infection can cause acute respiratory tract infection with atypical symptoms, including severe complications, in children.

[Detecting the occult HBV infection: a laboratory study].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Detection and clinical characterization of WU polyomavirus in acute respiratory tract infection in children].

OBJECTIVE: 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. METHOD: 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. RESULT: 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. CONCLUSION: 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.

[Preparation and selection of the monoclonal antibody used for kits to detect BNP32 which was applied to clinical research].

AIM: To prepare the monoclonal antibody (mAb) used for kits to detect the BNP32 antigen by means of double-antibody sandwich ELISA assay. Comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL. METHODS: BALB/c mice were immunized with purified recombinant BNP32 protein and by routine hybridoma technique, Then comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL. RESULTS: 16 hybridoma cell lines secreting potent mAb against BNP32 were obtained. The subtype of these 16 mAb were found to be IgG1, IgG2a and IgM, then their cross-blocking properties were analyzed, when they reacted with the BNP32 protein in direct ELISA in order to offer the valuable data for selecting feasible pair of mAb in the detection of the BNP32 antigen. All the mAbs were used for the detection of BNP32 by double-antibody sandwich ELISA. In addition, the mAbs were purified and HRP-labelled in advance. Finally, we had successfully screened a pair of mAbs, which exhibited a sensitivity of 20 ng/L for the detection of BNP32 antigen. The two detection methods have very good consistency (kappa=0.828>0.75)between double-antibody sandwich ELISA and Roche ECL. There was no statistics significance on differences between these two methods(P>0.05). CONCLUSION: It is evident that this pair of mAb shows excellent detection of BNP32. The double-antibody sandwich ELISA can be good apply to detect BNP level in clinical congestive heart failure patients.

[Viral pathogens of acute lower respiratory tract infection in hospitalized children from East Guangdong of China].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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%). CONCLUSIONS: 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.

[Detecting hepatitis B virus large surface protein to filtrate the occult HBV infection].

OBJECTIVE: Detecting hepatitis B virus large surface protein (HBLP) with serological method to filtrate the occult HBV infection and study the clinical detection strategy. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Detection of rpoB gene mutations in Mycobacterium tuberculosis].

ABSTRACT With Mycobacterium tuberculosis H37Rv as a control, the mutations of rpoB gene from 37 Mycobacterium tuberculosis clinical isolates (18 rifampin-resistant strains and 19 rifampin-susceptible strains) have been detected by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) under standard condition (no glycerol in gel). For the strains in which mutations had not been detected under standard condition, PCR-SSCP analysis was performed again on condition that 8% glycerol was added to gel. In addition, the PCR products of rpoB genes of 30 strains (18 rifampin-resistant strains and 12 rifampin-susceptible strains) were detected by DNA sequencing. After twice analyses of PCR-SSCP, in 18 rifampin-resistant strains, the patterns of 17 strains were found to be abnormal; and in 19 rifampin-susceptible strains, the patterns of 16 strains were found to be normal. Compared with routine drug susceptibility test, the sensitivity of PCR-SSCP 94.4% is higher for rifampin resistance detection, and specificity of PCR -SSCP 84% seems lower. The results of sequence analysis were shown as: amomg 18 rifampin-resistant strains, only one has deletion in codons 513 and 514 of rpoB gene, which is a new report, the others have point mutation; among 12 rifampin- susceptible strains, 3 positive strains in SSCP occur gene mutations directly related to rifampin resistance. Then compared with DNA sequencing, the accuracy, sensitivity and specificity of PCR-SSCP are 96.7%, 95.2% and 100% respectively. Therefore, the sensitivity of mutation detection in SSCP could be improved by adding glycerol in gel. It is feasible and efficient to detect the mutation of rpoB gene in Mycobacterium tuberculosis by PCR-SSCP. This method is valuable to evaluate the rifampin resistance of Mycobacterium tuberculosis in clinical application for curing tuberculosis.