ABSTRACT
Objective:To analyze the changes in T lymphocyte subsets, B lymphocytes and NK cells in children with active tuberculosis (TB) and their clinical significance.Methods:T lymphocyte subsets, B lymphocytes and NK cells in peripheral blood samples of 106 patients with acute TB (TB group) and 106 healthy children (healthy control group) were detected by flow cytometry and compared between different groups.Results:The percentages of CD3 + T, CD4 + T and NK cells as well as the CD4 +/CD8 + T cell ratio were significantly lower in the TB group than in the healthy control group ( Z=-3.783, P=0.000; Z=-5.401, P=0.000; Z=-3.434, P=0.001; Z=-2.014, P=0.044). The percentages of double negative T (DNT) and B cells in the TB group were significantly higher than those in the healthy control group ( Z=2.765, P=0.006; Z=6.880, P=0.000). No significant difference in the percentage of CD8 + T or double positive T (DPT) cells was observed between the two groups ( P>0.05). The expression of peripheral lymphocyte subsets varied in TB children of different age groups (0-<3, 3-<6, 6-<10 and 10-<16 years old). There were significant differences in CD3 + T, DNT and B cells among the four age groups ( H=10.081, P=0.018; H=14.583, P=0.002; H=8.498, P=0.037). The percentage of CD4 + T cells was significantly lower in children with extrapulmonary TB than in those with pulmonary TB ( Z=-3.068, P=0.002). No statistically significant difference in other lymphocyte subsets was found between children with extrapulmonary and pulmonary TB ( P>0.05). Conclusions:Tuberculosis could lead to immune dysfunction in children. Dynamic monitoring of the changes in peripheral lymphocyte subsets in children with TB could be conducive to better assessment of immune status and providing personalized treatment.
ABSTRACT
Objective:To analyze the epidemiological characteristics of respiratory syncytial virus (RSV)-caused respiratory diseases in children in Hebei Province and the changes in peripheral blood lymphocyte subsets and inflammatory indexes.Methods:A total of 9 491 sputum specimens and 9 491 paired peripheral blood specimens were collected from children admitted to Hebei Children′s Hospital for respiratory infection in 2018. RSV-positive sputum specimens were screened by multiple detection reagents for 13 kinds of respiratory pathogens. Flow cytometry was uses to detect T and B lymphocytes and NK cells in peripheral blood samples of randomly screened RSV-positive children. Procalcitonin (PCT) was measured by Roche E411 luminescence analyzer. Hypersensitive C-reactive protein (hs-CRP) was detected by Roche Cobas 8000 C701 biochemical analyzer. White blood cells (WBC) were measured by Sysmex XN-BN3 hematometer.Results:The positive rate of RSV in children with respiratory diseases was 13.08% in Hebei Province in 2018. There were significant differences in RSV-positive rates among different age groups (χ 2=479.297 6, P<0.000 1). The positive rate of RSV decreased gradually with age (χ 2=-20.282 7, P<0.000 1) and was higher in male than in female (χ 2=34.552 7, P<0.000 1). The incidence of co-infection of RSV with other respiratory pathogens was 29.49% (366/1 241), mainly caused by human rhinovirus (HRV, 150/1 241) and adenovirus (ADV, 40/1 241). The main epidemic seasons of RSV infection were winter and spring. The epidemic trends of simple RSV infection and co-infection were consistent. There were significant differences in inflammatory indexes, WBC ( P<0.01), CD4 + ( P=0.015) and CD4 + /CD8 + cells ( P=0.016) between simple RSV infection and co-infection groups. Conclusions:RSV was a common pathogen causing respiratory diseases in children in Hebei Province. The younger the children were, the more likely they were to be infected with RSV. RSV infection was easily complicated by HRV or ADV infection. The epidemic seasons of RSV infection in Hebei were winter and spring. Both simple infection and co-infection of RSV might result in immune dysfunction.
ABSTRACT
In recent years, digital PCR (dPCR), as a novel nucleic acid amplification detection technique, contrasts real-time fluorescent quantitative PCR (qPCR) without the need to establish a standard curve, "single molecule template amplification" absolute quantification, insensitive to inhibitors that affect PCR efficiency, and excellent sensitivity, specificity, extremely strong repeatability and the development of the existing commercial platform technology that has great application advantages in virology and nucleic acid quantification. In this review, we address the application advantages of digital PCR in virus detection and describe its future development.
ABSTRACT
Objective@#To systematically review the diagnostic accuracies of multiplex real time polymerase chain reaction (MRT-PCR) technique for detection of respiratory syncytial virus (RSV) and adenovirus (ADV).@*Methods@#PubMed, EMBASE, Cochrane, Wanfang and CNKI databases were searched from January1 2010 to January1 2018, to collect reports on MRT-PCR for detection of common respiratory viruses. Then two authors independently exacted the data and assessed the risk of bias of included studies by using the QUADAS-2 tool. Meta-disc 1.4.@*Results@#Ten articles with 2528 cases were eligible for analysis. The result of meta-analysis showed that, the pooled Sen, Spe and area under SROC curve, for detecting RSV were 0.87 (95% CI 0.83 to 0.90), 0.98 (95% CI 0.97 to 0.98) and 1.00. The pooled Sen, Spe, and area under SROC curve of MRT-PCR for detecting ADV were 0.64 (95% CI 0.56 to 0.71), 0.99 (95% CI 0.98 to 0.99) and 0.99. Deeks test indicated that no publication bias was found.@*Conclusions@#The sensitivity of MRT-PCR in RSV and ADV detection is still to be improved, but the overall detection ability is good which deserves to be recommended for clinical use.
ABSTRACT
Objective To analyze the molecular epidemiology of adenovirus ( ADV) causing acute respiratory diseases and to investigate the mixed infection of ADV and other respiratory pathogens in Hebei Province. Methods Sputum samples were collected from inpatient children with acute respiratory diseases at Children′s Hospital of Hebei Province between June 2017 and May 2018. Multiplex reverse transcription PCR assay was used to detecte 13 kinds of respiratory pathogens. Nested PCR was performed to amplify ADV hexon gene and the amplified products were then sequenced. Results A total of 353 ADV-positive speci-mens were detected in 8839 specimens with a positive rate of 3. 99％. Significant difference in the positive rate of ADV was not observed between male and female patients (χ2=0. 0003, P=0. 99), but found among different age groups (χ2=115. 69, P<0. 001). All isolated ADV strains belonged to 11 serotypes, which were type 1 (16. 15％, 57/353), type 2 (35. 98％, 127/353), type 3 (21. 25％, 75/353), type 4 (1. 13％, 4/353), type 5 (11. 33％, 40/353), type 6 (3. 97％, 14/353), type 7 (8. 22％, 29/353), type 31 (0. 28％, 1/353), type 41 (0. 28％, 1/353), type 55 (0. 28％, 1/353) and type 57 (1. 13％, 4/353). Among the 353 ADV-positive specimens, 259 were mixed infections mainly caused by ADV and human rhinovirus (35. 52％). ADV and respiratory syncytial virus co-infections accounted for 12. 74％ and 33. 20％ of the mixed infections involved three or more pathogens. ADV could be detected throughout the year, especially in September and April to May. The predominant serotypes were types 1, 2 and 3. The av-erage ages of the two groups of ADV infection alone and ADV mixed infection were (27. 56±24. 67) months and (21. 33 ±20. 28) months, respectively, and the difference between them was statistically significant (P=0. 037). The incidence of ADV 2 infection alone was 25. 77％ (25/97), which was lower than that of ADV 2-involved mixed infection [39. 84％ (102/256),χ2=6. 05, P=0. 014]. However, the rate of ADV 7 infection alone was significantly higher than that of ADV 7-involved mixed infection [16. 49％ 16/97) vs 5. 08％ (13/256),χ2=6. 05, P<0. 001]. Conclusion ADV 1, ADV 2 and ADV 3 were the predominant serotypes circulating in Hebei Province from June 2017 to May 2018, especially in September and April to May. The younger the patients were, the higher the incidence would be. ADV 2 was prone to cause mixed infections with other respiratory pathogens, while ADV 7 was less common in mixed infections. Younger pa-tients were more susceptible to mixed infections. The most common co-infection was caused by ADV and hu-man rhinovirus.
ABSTRACT
Objective To develop a tetra-primer amplification refractory mutation system PCR(T-ARMS-PCR)assay for detecting four single nucleotide polymorphisms(SNPs): rs1801133,rs1801131, rs1805087 and rs1801394 associated with folate metabolism in a single reaction tube.Methods Methodology was developed.Then, it applied the established method to analyze 150 physical examination children′s anticoagulant blood samples admitted to the Department of Clinical Laboratory, Children′s Hospital of Hebei Province from January 2017 to April 2017.Four sets of chimeric primers consisting of a universal Tag sequence and targeting rs 1801133,rs1801131,rs1805087 and rs1801394 fused to the specific sequence were designed according to the T-ARMS-PCR principle and the multiplex chimeric primers strategy.A single tube PCR was then conducted after optimizing the primer concentrations and the reaction conditions.The amplified products were analyzed by QIAxcel capillary electrophoresis, and the corresponding SNP genotypes of 150 samples were identified.Furthermore,all samples were verified by direct sequencing.And the Hardy-Weinberg Equilibrium(HWE)testing of four SNPs from 150 samples were conducted by SPSS17.0 Chi-square test.Results The improved T-ARMS-PCR combined with capillary electrophoresis can accurately verify eight different alleles of the four SNPs associated with folate metabolism at one time in 3 hours.Four SNPs of 150 whole blood samples were accurately classified and the results were completely consistent with direct sequencing.All the genotype frequencies of these four SNPs were in HWE (χ2rs1801133=0.69, Prs1801133=0.40; χ2rs1801131=0.21, Prs1801131=0.64; χ2rs1805087=3.32, Prs1805087=0.07;χ2rs1801394=1.91, Prs1801394=0.17).Conclusions The proposed improved T-ARMS-PCR combined with capillary electrophoresis in this study can accurately verify four SNPs associated with folate metabolism in a single reaction tube.This method might be a valuable tool to specifically guide the folate supplement in general population.
ABSTRACT
Objective To analyze the levels and clinical significances of IL-18,Caspase-3 and nerve tissue-specific protein S-100B at different disease extent and different stages of infants with hypoxicischemic encephalopathy (HIE).Methods This study was clinical experimental studies.Sixty-seven infants with HIE (23 cases of mild HIE,23 cases of moderate HIE,21 cases of severe HIE) from February 2008 to June 2009 in Hebei Children's Hospital were enrolled.The levels of IL-18,Caspase-3 and S-100B protein in all samples were measured at acute phase (1 d,3 d) and recovery phase (7 d) by ELISA method.Twenty healthy full-term neonates were selected as the normal control group.Multi-factor analysis of variance and Pearson correlation test was used for statistical analysis.Results The levels of the three indicators in the moderate and severe group were higher than the normal control group.In the moderate group,IL-18 levels were(132.15 ± 9.87),(150.31 ± 15.04) and (87.91 ± 9.93) ng/L,Caspase-3 levels were (5.79 ±0.64),(7.36 ± 1.57)and (3.79 ±0.61) μg/L,S-100B levels were(6.82 ±0.61),(9.62 ± 1.29) and (10.76 ± 1.64) μg/L.In the severe group,IL-18 levels were (160.23 ± 16.03),(189.86 ± 18.32) and (107.35 ± 13.02) ng/L;Caspase-3 levels were (6.86 ± 1.02),(9.54 ± 1.43) and (5.25 ± 0.71) μg/L;S-100B levels were(8.90 ± 0.32),(12.54 ± 0.89)and(13.53 ± 0.75) μg/L.In the normal control group,IL-18 levels were (71.08 ± 11.52),(72.53 ± 11.05) and (71.93 ± 11.30) ng/L; Caspase-3 levels were (2.84 ± 0.52),(2.98 ± 0.53) and (2.87 ± 0.52) μg/L; S-100B levels were (1.50 ± 0.25),(1.62 ±0.30)and(1.53 ±0.29) μg/L IL-18 levels,Caspase-3 levels and S-100B levels in severe group were higher than the moderate group and the mild group were higher than the mild group.IL-18 levels were (73.46 ± 4.77),(77.59 ± 4.02) and (72.87 ± 6.92) ng/L ; Caspase-3 levels were (3.13 ± 0.31),(3.63±0.40) and (3.26 ±0.45) μg/L;S-100B levels were(3.68 ±0.40),(5.851 ±0.63) and(6.95 ± 0.58) μg/L in the mild group.S-100B levels in the mild group were higher than that in the normal control group.The IL-18 and Caspase-3 levels were risen in the third day to the first day in the acute phase of the moderate group and severe group,decreased in the recovery phase.Serum S-100B protein levels in the acute and recovery phase increased gradually,and there was no correlation between the three indicators (r-=0.321,0.14,0.48,P=0.438,0.974,0.911 respectively).Conclusions IL-18,Caspase-3 and S-100Bwere involved in the pathophysiological process of HIE.The levels were closely related to the severity and disease progression of HIE,the severer of the illness,and the higher of the levels.Dynamic monitoring the changes of the three indicators may contribute to an early diagnosis,condition monitoring and prognosis of HIE.