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Objective:To evaluate the clinical outcomes of patients with borderline developmental dysplasia of the hip (BDDH) and cam-type femoroacetabular impingement syndrome (FAIS) after hip arthroscopy.Methods:Data were retrospectively reviewed for patients with BDDH and cam-type FAIS who underwent hip arthroscopy surgery from June 2017 to December 2019. A total of 32 patients were enrolled, with a mean age of 36.13±8.67 years (range, 20-50 years), including 15 males and 17 females. The preoperative lateral center-edge angle was 22.3°±1.6° (range 20.1°-24.7°), while the preoperative α angle was 64.1°±4.6° (range, 56.0°-69.8°). All patients were treated with arthroscopic limited acetabular plasty, labral repair, femoral osteoplasty, and capsular plication after excluding from external hip diseases by ultrasound-guided hip blocking test. The visual analogue scale (VAS), modified Harris Hip Scores (mHHS) and International Hip Outcome Tool-12 (iHOT-12) scores were used to evaluate the clinical effects.Results:All patients were followed up, and the mean follow-up time was 2.5±0.8 years (range, 2.0-4.7 years). The VAS score decreased from 6.07±1.56 to 1.96±0.92 at 1 year and to 1.86±1.01 at 2 years after operation ( F=112.64, P<0.001); the mHHS score increased from 53.87±13.04 to 86.12±8.64 at 1 year and to 88.71±8.15 at 2 years after operation ( F=101.70, P<0.001); the iHOT-12 score was improved from 40.00±7.33 to 76.27±9.50 at 1 year and to 78.67±10.31 at 2 years after operation ( F=134.91, P<0.001). The α angle improved to 40.27°±4.52° (range, 34.8°-49.7°) with significant difference ( t=9.24, P<0.001). Conclusion:Hip arthroscopy can achieve satisfied short-term outcomes in treating BDDH and cam-type FAIS with few complications and less trauma.
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Objective:To explore the performance of the commonly used whole blood C-reactive protein (CRP) detection systems and give related recommendation on the performance requirements of detection systems.Methods:A total of 7 540 venous blood samples from 26 maternal, child and children′s hospitals were collected to conduct this multi-center study on the analytical performance of 5 commonly used whole blood CRP detection systems from March to April in 2019. The blank check, carryover, repeatability, intermediate precision, linearity, sample stability, influence of hematocrit/triglyceride/bilirubin, comparison with SIEMENS specific protein analyzer and trueness were evaluated. The 5 systems included BC-5390CRP autohematology analyzer, AstepPLUS specific protein analyzer, Ottoman-1000 Automated Specific Protein POCT Workstation, i-CHROMA Immunofluorometer equipment Reader and Orion QuikRead go detecting instrument. The 5 systems were labeled as a, b, c, d and e randomly.Results:Within the 5 systems, all values of blank check were less than 1.00 mg/L, the carryovers were lower than 1.00%. The repeatability of different ranges of CRP concentrations including 3.00-10.00, 10.00-30.00 and>30.00 mg/L were less than 10.00%, 6.00% and 5.00%, respectively, and the intermediate precision was less than 10.00%. The linearity correlation coefficients of the 5 systems were all above 0.975, while the slope was within 0.950-1.050. Whole blood samples were stable within 72 hours both at room temperature (18-25 ℃) and refrigerated temperature (2-8 ℃). The CRP results were rarely influenced by high triglyceride or bilirubin, except for the immmunoturbidimetric test based on microparticles coated with anti-human CRP F(ab) 2 fragments. When triglyceride was less than 15.46 mmol/L, the deviation of CRP was less than 10.00%. When bilirubin was less than 345.47 μmol/L, the deviation of CRP was less than 10.00%. CRP was more susceptible to Hct on the systems without Hct correction. The deviation of CRP between different Hct dilution concentration and 40% dilution concentration can reach as high as 67.48%. The correlation coefficients ( r) of 5 systems were all more than 0.975 in the range of 0-300.00 mg/L compared with Siemens specific protein analyzer. All systems passed the trueness verification using the samples with specified values of 12.89 and 30.60 mg/L. Conclusion:The performance of 5 systems can basically meet the clinical needs, but it is suggested that the whole blood CRP detection system without automatic Hct correction should be modified manually.
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Objective To investigate the prevalence of common enterovirus infections among children in Hangzhou ,2016 .Methods A total of 2977 of throat swabs samples or stool samples from suspected children with enterovirus infection at Hangzhou Children′s Hospital in 2016 were collected . Enterovirus universal nucleic acid ,enterovirus type 71 (EV71) ,Coxsackievirus A (CoxA)16 ,CoxA6 , CoxA4 and CoxA10 were detected by real-time fluorescent quantitative polyoneras chain reaction .The detection positive rates among children with different genders and ages were compared by χ2 test .Results The total enterovirus positive rate of 2977 specimens was 49 .7％ (1480/2977) .Among them ,CoxA6 , EV71 ,CoxA16 ,CoxA4 ,CoxA10 and other enteroviruses accounted for 30 .2％ (447) ,19％ (281) , 10 .2％ (151) ,6 .8％ (101) ,3 .4％ (51) and 30 .3％ (449) ,respectively .Among 1480 enterovirus positive children ,882 cases were male and 598 were female ,with no statistical significance (χ2 = 4 .564 , P=0 .471) .The detection rates of enterovirus in children with hand ,foot and mouth disease (HFMD ) , herpangina and other diseases were 64 .4％ (1051/1632) ,52 .4％ (323/616) and 14 .5％ (106/729) , respectively .The difference among groups was significant (χ2 =503 .387 , P<0 .01) .The prevalences of CoxA6 ,EV71 and CoxA16 in enterovirus-positive HFMD children were 36 .7％ (386) ,25 .3％ (266) and 12 .4％ (130 ) , respectively . The prevalences of CoxA4 , CoxA6 and CoxA10 in enterovirus-positive herpangina children were 20 .1％ (65 ) , 14 .9％ (48 ) and 8 .7％ (28) , respectively , and those of entrovirus ,CoxA4 and CoxA6 in other enterovirus-positive specimens were 59 .4％ (63) ,14 .2％ (15) and 12 .3％ (13 ) , respectively . The enterovirus constituent ratios among children with HFMD , herpangina and other diseases was statistically different (χ2 =399 .758 ,P<0 .01) .The positive rates of enterovirus among different age groups were statistically different (χ2 = 142 .899 , P< 0 .01 );the constituent ratios of enterovirus among different age groups were also statistically different (χ2=106 .160 , P<0 .01) .Conclusion The enterovirus detection rate of CoxA6 is highest ,followed by EV71 and CoxA16 among children in Hangzhou ,2016 .
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Objective@#To investigate the etiology composition of enterovirus (EV) in patients with severe hand, foot, and mouth disease (HFMD) in children. To assess the diagnostic value of cerebrospinal fluid (CSF) tests in severe HFMD, and to find the key laboratory tests for severe HFMD.@*Methods@#A total of 288 hospitalized cases of children clinically diagnosed with severe HFMD in Hangzhou Children′s Hospital were included from March to July 2016. Throat swabs were collected and enterovirus nucleic acids were detected by fluorescence quantitative reverse transcription (RT)-PCR. Synchronous CSF and serum samples were collected for EV-A71 and CV-sackievirus A16 (CV-A16)-IgM antibody detection. CSF samples underwent routine and biochemical tests. Normally distributed continuous variables were compared using t test. Non-normal distribution continuous variables were compared using Mann-Whitney U test. Differences between categorical variables were compared with χ2 test.@*Results@#The total positive rate of enterovirus nucleic acid EV-A71/CV-A16/EV by fluorescence quantitative RT-PCR in the 288 cases of children clinically diagnosed with HFMD was 83.7% (241/288), including EV-A71 55.2% (159/288), CV-A16 4.9% (14/288) and the other enterovirus 23.6% (68/288). Among the other enterovirus group, there were 29.4% CV-A6 (20/68) , 16.2% CV-A4 (11/68) and CV-A10 10.3% (7/68). The total positive rate of combined serum and CSF detection of EV nucleic acid and EV-A71 and CV-A16 IgM antibody was 98.3% (283/288). EV nucleic acid positive rate was 83.7% (24/288). The positive rates were statistically different (χ2 =37.289, P=0.000). The CSF nucleated cells count in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-4.472 and -9.991, respectively, both P<0.05). The CSF nucleated cells positive rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ2=43.857 and 133.078, respectively, both P<0.05). The CSF protein level in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-3.151 and -5.255, respectively, both P<0.05). The CSF protein positive (>400 mg/L) rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ2=4.956 and 11.795, respectively, both P<0.05). The CSF nucleated cell counts and positive rates in EV-A71 IgM antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The CSF protein level and elevated proportion in antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The lactate dehydrogenase concentration in antibody-positive subgroup was significantly higher than those in antibody-negative subgroup (P<0.05). The EV-A71 IgM antibody in serum was significantly correlated with the antibody in CSF (r=0.600, P<0.05).@*Conclusions@#EV-A71 is still the most important pathogen of severe HFMD in Hangzhou in 2016. Other enterovirus such as CV-A6, CV-A4 increases compared to those in 2014 and 2015. The CSF routine and biochemical tests and the IgM antibody levels can serve as an important indicator for the diagnosis of children with severe HFMD.
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Objective@#To observe the intestinal viral shedding time in patients with hand, food and mouth disease (HFMD) induced by coxsackievirus A6 (CA6).@*Method@#Throat swab specimens and stool specimens of HFMD children were collected from those admitted to Hangzhou Children′s Hospital between May and October 2015, while fluorescence quantitative PCR was used to detect the viral load.Eeighteen cases of HFMD children were followed up, who were confirmed as CA6 infection via laboratory tests.Stool specimen was collected every 4-7 days, and fluorescence PCR was used for virus nucleic acid detection until the stool viral nucleic acids of infected children turned to be negative.The intestinal virus shedding time of CA6-infected HFMD was compared with the intestinal virus shedding time of 65 children with enterovirus 71 (EV71) infection and 44 children with coxsackievirus A16 (CA16) infection of the previous studies (from May to September 2012).@*Result@#The median stool viral load was 25×105 copies/ml (55×104 copies/mL, 9×106 copies/ml) in CA6-infected children.The numbers of stool virus nucleic acid turning negative were 0 case, 4 cases, 9 cases, 3 cases and 2 cases in 18 children at 1st, 2nd, 3rd, 4th, 5th weeks. At 5th week, the stool virus nucleic acid of children in CA6 group all turned to be negative.The positive rates of stool virus nucleic acid in EV71 group and CA16 group at the 5th week, however, were 31% and 27% respectively.There were statistically significant differences in distribution of positive rate of stool virus nucleic acid between CA6 infected children with EV71 and CA16 infected children (χ2=13.894, 10.698, P<0.05).@*Conclusion@#The longest intestinal virus shedding time for CA6-infected HFMD children was 5 weeks, which is obviously shorter than that of EV71- infected children and CA16-infected children.
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Objective To explore the clinical application value of serum amyloid A (SAA),C-reactive protein (CRP) and SAA/CRP in early diagnosis of hand-foot-mouth disease (HFMD).Methods The serum levels of SAA and CRP were detected in 873 children with HFMD who were admitted in Hangzhou Children's Hospital from April to December in 2015.And 487 healthy children were enrolled as healthy control group.SAA was measured by nephelometry assay,and CRP was measured by immunoturbidimetry.Variables were compared using Mann-Whitney U test and diagnostic value was measured by using receiver operating characteristic (ROC) curve.Results The median levels of SAA in HFMD group and control group were 330.5 and 4.0 mg/L,respectively,which was statistically different (Z=-29.02,P＜0.01);the median levels of CRP were 10.0 and 1.0 mg/L,respectively,with statistical difference (Z =23.79,P＜ 0.01);and the median SAA/CRP were 23.06 and 2.40,respectively,with statistical difference (Z=-24.79,P＜0.01).In ROC curve comparison,the area under the curve of SAA (0.980) was higher than those of SAA/CRP ratio (0.911) and CRP (0.899) for diagnosing HFMD.When using 10.30 mg/L as the cut off value for SAA according to the ROC curve,the sensitivity was 91.6％ and the specificity was 96.6％.Conclusions Joint detection of SAA and CRP can improve the efficiency in the early diagnosis of HFMD.SAA can provide useful auxiliary information for the diagnosis of HFMD,which is worthy of extensive clinical application.
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<p><b>OBJECTIVE</b>To detect the anti-enterovirus 71 (EV71) IgM level in cerebrospinal fluid (CSF) of children with severe hand, foot and mouth disease (HFMD) induced by EV71 and then analyze the relationships among the IgM antibody levels, CSF routine examination and patients' clinical features, and thus to evaluate the clinical significance of anti-EV71 IgM as a new indicator for early diagnosis of children with severe HFMD induced by EV71.</p><p><b>METHOD</b>A total of 294 laboratory-confirmed cases of children with severe HFMD infected with EV71 were enrolled into the research group from March 2014 to June 2014, consisting of 53 fatal cases and 241 severe cases, and their CSF samples underwent enzyme-linked immunosorbent assay (ELISA) for anti-EV71 IgM levels, CSF routine and biochemical tests. Forty-one cases of children with severe HFMD induced by other enteroviruses were collected as antibody-testing control group during the same period.</p><p><b>RESULT</b>In the research group, the total positive rate of anti-EV71 IgM in 294 CSF samples of children with severe HFMD infected by EV71 was 60.2% (177/294); the positive rate of anti-EV71 IgM in the fatal HFMD subgroup was 62.3% (33/53); the positive rate of anti-EV71 IgM in the severe HFMD subgroup was 59.8% (144/241). In the control group, the results of CSF anti-EV71 IgM tests were all negative (0/41). In the research group, patients in antibody-positive subgroup (2.5±1.2) years old were younger than those in antibody-negative subgroup (2.9±1.1) years old (t=2.595, P=0.010). And within the antibody-positive subgroup, the patients ((1.9±0.7) years old) with fatal type disease were younger than those ((2.6±1.2) years old) with severe type disease (t=3.150, P=0.002). The CSF nucleated cells count and positive rates (105 (56,180) ×10(6) /L; 97.7% (173/177)) in antibody-positive subgroup were higher than those (62(30,150) ×10(6) /L; 83.8% (98/117)) in antibody-negative subgroup (Z=3.663, P=0.000; χ(2)=19.089, P=0.000). In antibody-positive subgroup, the percentage of monocytes (57±25)% was higher than that of polykaryocytes (43±25)%. In antibody-negative subgroup, the percentage of monocytes (50±26)% was close to that of polykaryocytes (50±26)%. In the antibody-positive subgroup, the ratio of the patients with nucleated cells count higher than 100×10(6)/L in fatal type group and severe type group was 69.7% (23/33) and 47.2% (68/144) respectively (χ(2)=5.429, P=0.02). The CSF protein quantity and positive rates in antibody-positive subgroup were higher than those in antibody-negative subgroup (Z=2.158, P=0.031; χ(2)=5.921, P=0.015).</p><p><b>CONCLUSIONS</b>The anti-EV71 IgM levels in CSF can serve as an important indicator for early diagnosis of children with severe HFMD induced by EV71. And the anti-EV71 IgM levels in CSF correlated to the CSF nucleated cells count and classification and CSF protein quantity. In the antibody-positive subgroup, the higher the nucleated cell count or the younger the age, the higher the possibility of patients to develop into fatal cases.</p>
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Objective To assess the value of combined detection of enterovirus nucleic acid and antibody in early etiological diagnosis for hand-foot-and-mouth disease ( HFMD).Methods A case-control study was conducted.A total of 1 066 cases of children clinically diagnosed with HFMD from Hangzhou Children′s Hospital were involved into the research group from January to June 2014, consisting of 401 common cases and 665 severe cases; Throat swabs and serum samples from these children underwent combined detection for EV71/CA16/EV of enterovirus nucleic acid by fluorescence quantitative RT-PCR and for EV71/CA16-IgM by ELISA.All data were analyzed with SPSS 16.0.Results The total positive rate of enterovirus nucleic acid EV71/CA16/EV by fluorescence quantitative RT-PCR in the 1 066 cases of children clinically diagnosed with HFMD was 75.52%( 805/1 066 ) ( 95%CI: 72.80%-78.05%).But the total positive rate of combined detection was 91.46%( 975/1 066 ) ( 95%CI:89%.58-93.04%).The total positive rate of combined detection is higher than that of RT-PCR test(χ2 =98.338,P=0.000).The positive rate of EV71 type of combined detection was 64.63%(689/1 066)(95%CI:61.67%-67.49%),which is 15.38%higher than that of RT-PCR test 49.25%(525/1 066)(95%CI:46.21%-52.29%)(χ2 =51.453, P=0.000).In 665 severe cases of HFMD, the total positive rate of combined detection was 96.69%(643/665)(95%CI:94.95%-97.87%), which is higher than that of RT-PCR test 79.25%(527/665)(95%CI:75.92%-82.22%)(χ2 =95.607, P =0.000).In the severe cases, the positive rate of EV71 type of combined detection was 87.52%( 582/665 ) ( 95%CI:84.71%-89.89%) , which is 18.95% higher than that of RT-PCR test 68.57%(456/665) (95%CI:64.87%-72.06%) (χ2 =69.665, P=0.000).In the fatal cases, the positive rate of EV71 type of combined detection was 95.92%(94/98) (95%CI:89.28%-98.68%).Conclusions The combined detection of enterovirus nucleic acid and specific IgM antibody can significantly increase the positive rate of HFMD, especially for severe cases.The combine detection increases both the total positive rate and EV71 positive rate.Thus it has a high potential for becoming a new guidelines for laboratory diagnosis of HFMD.
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Objective To investigate cerebrospinal fluid characteristics and clinical features in children with severe hand,foot and mouth disease (HFMD) induced by enterovirus 71 (EV71) infection.Methods A total of 114 children with severe HFMD,in whom EV71 was detected by reverse transcription polymerase chain reaction (RT-PCR),were admitted in Hangzhou Children's Hospital during May and August 2013.Seventy-eight children with severe HFMD induced by other enteroviruses admitted at the same period served as controls.The results of cerebrospinal fluids (CSF) routine examination and biochemical tests,and the clinical symptoms were compared between two groups.Differences in enumeration data were compared with x2 test,and measurement data were compared with Mann-Whitney U test.Results The incidences of vomiting and limb shaking in EV71 infection group were 35.1％ and 50.9％,which were higher than those in control group (x2 =7.864 and 19.682,P ＜ 0.05).The incidence of limb shaking in children with nucleated cells count ≥ 100 × 106/L in EV71 group was higher than that with nucleated cells count ＜ 100 × 106/L (72.3％ vs.35.8％,x2 =14.740,P =0.000).The nucleated cells count,protein quantity and their positive rates in EVT1 infected group were higher than those in control group (Z =-9.458 and-6.591,P=0.000; x2=105.421 and 10.932,P =0.000 and 0.001).Conclusion The symptoms of nervous system damage and abnormal CSF examination were more serious in HFMD induced by EV71 infection,and in EV71 infected patients the incidence of limb shaking is correlated with nucleated cell count in CSF.
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Objective To establish a rapid and high sensitive assay of real-time fluorescence loop-mediated isothermal amplification assay for clinical detection of HIV-1 DNA.Methods Four loop primers were constructed for loop-mediated isothermal amplification ( LAMP) assay based on six conserved regions selected from HIV gene sequence .SYBR Green Ⅰdye was added into the established LAMP assay and its specificity and sensitivity were evaluated .Results The real-time fluorescence LAMP assay for the detection of HIV-1 DNA was successfully established .It was used for the detection of HIV-1 DNA among 200 patients with HIV infection, of which 195 cases were positive.Moreover, 50 healthy subjects were found HIV-1 DNA negative tested by the real-time fluorescence LAMP assay .Quantitative testing for HIV-1 DNA showed that the lowest and the highest detectable amount were 51 copies/ml and 8.21×106 copies/ml respectively, and the average amount was 5.78×105 copies/ml.HIV viral loads ranging from 1×105 to 10×105 were detected in 162 of 200 patients (83.08%).The ten times dilution method showed that the lowest detection limit of the assay was 50 copies/ml.The crossover experiment indicated that the specificity of the assay was 100%as none of HBV-DNA, HSV-DNA and HCV-RNA was determined by the assay .Conclusion The present study shows that 97.5%of the patients with HIV infection are confirmed HIV-1 DNA positive by the real-time fluorescence LAMP assay , suggesting that the real-time fluorescence LAMP assay is a rapid and sensi-tive assay with high specificity and could be applied for clinical detection of HIV-1 DNA.
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Objective To develop a method of simultaneous detection and Gram classification for pathogens causing sepsis with gram-specific probes based real-time PCR. Methods A pair of universal primers and a set of probes including Gram-positive probe and Gram-negative probe were designed based on the bacterial highly conserved region of 16SrRNA gene. With the gram-specific probes based real-time PCR, 35 clinical frequently-isolated strains including 17 gram-positive and 18 gram-negative bacteria were identified correctly with the corresponding gram probe. The blood samples from 512 cases of suspected septicemia, who were hospitalized in our neonatal ward and the NICU and developed clinical signs suggestive of infection, were tested with routine culture and bacterial gram-specific probes based real-time PCR separately. Results The detection limit of the gram-specific probes based real-time PCR assay was 10 CFU of the bacteria. The 35 isolates could be detected and classified correctly by gram-specific probes based real-time PCR. The PCR results were all negative for Cytomegalo virus, EB virus, hepatitis B virus, Cryptococcns neoformans, candida albican, human genomic DNA and negative control. The gram-specific probes based real-time PCR appeared to be quite specific. For 512 blood specimens from the patients with suspicious neonatal sepsis, the positive rate of the gram-specific probes based real-time PCR array was 8.20% (42/512,), which is significantly higher than that of blood culture (32/512, 6.25%) (χ2=8.10, P<0.01). When blood culture was used as a standard, the sensitivity of the gram-specific probes based real-time PCR was 100%. The specificity was 97.92% and the accuracy was 98.05%. Canclusions Cram-specific probes based real-time PCR with universal primers and gram-specific probes are developed. This study suggests that the bacterial gram-specific probes based real-time PCR are very useful for the rapid and accurate diagnosis of bacterial infection.
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Objective To study the current situation of reproductive tract infection among bearing age mar-fled women in countryside and influence factors. Methods We had examined 4916 women. Questionnaire investiga-tion,gynecologic examination,the inspection of pathogen and the B-ultrasound were collected by face to face in the clinic. Results The general prevalence rate of RTI was 53.93%. The prevalence rates of pelvic infection and cervici-tis were 3.97% and 51.12%. The prevalence rates of germs, trichomonal and candiclal vaginitis were 12.51%, 3.60% and 7.71% respectively. The infection rate for only one,two or three kind of RTI were 33.08% ,19.73% and 1.08% respectively. The influence factors were : age, education, family economy, sanitary habits, graviclity, frequent in-tercourse,induced abortion and knowledge towards RTI. Conclusion Among bearing age married women in country-side province the prevalence rate of RTI was higher and also showed evidence of some influence factors.
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Objective To establish a method for rapid diagnosis and identification of seven species human herpesviruses infection.Methods Primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase genes in human herpesviruses, namely herpes simplex virus type 1 (HSV-1),HSV-2,varicella-zoster virus (VZV),Epstein-Barr virus (EBV),cytomegalovirus (CMV),and human herpes virus 6 (HHV-6A/6B).DNA microarrays were made by printing the oligonucleotide probes on the special glass slides.A total of 282 blood specimens from children with suspected infection were analyzed by this DNA microarray technique,and the results were compared with those of TaqMan PCR.Results The products of the seven human herpesviruses after PCR amplification could be used to identify the virus species with DNA microarrays.The detection limits were 10 copies/?l for HSV-1,HSV-2,VZV,EBV,CMV,HHV-6A,and HHV-6B,respectively.The assay did not show cross-reaction to the DNA extract of hepatitis B virus,staphylococcus aureus,E.coli,Candia albicans and human genome.Among the 282 samples,59 were positive for human herpesviruses DNA.Compared with those of TaqMan PCR,the sensitivity and specificity of the microarray assay were 96.7% and 99.5%,respectively,and the index of accurate diagnosis was 0.962.Conclusions This DNA microarray for identifying human herpesviruses species is specific and sensitive,and may serve as an efficient technique for simultaneous detection and species identification of human herpesviruses in clinical specimen.
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Objective To improve the speed and accuracy of bacteria detection, and develop the test of 16S rRNA genes PCR amplification plus gene chip hybridization to diagnose neonatal sepsis. Methods Bacterial 16S rRNA genes were detected in blood and CSF samples of 125 suspected neonatal sepsis, and the results were compared with blood culture, CSF culture, and non-specific diagnostic parameters (WBC, PLT, CRP). 30 non-infectious neonates were regarded as the negative control group. Gene chip test were performed by extraction of DNA, primers and probes design, PCR amplification, preparation of gene chip, hybridization, laser scan and reading of the results. 18 specific probes, including universal 1, universal 2, Gram positive bacterial probe, Gram negative bacterial probe 1, Gram negative bacterial probe 2, Staphylococcus aureus, Staphylococcus epidermidis, CoNS (Coagulase Negative Staphylococcus), Escherichia coli, Haemophilus influenzae, Listeria monocytogenes, Streptococcus pneumoniae, Streptococcus agalactiae, Bacteroides fragilis, Bacillus, Meningococcus, Corynebacterium, Propionibacterium, were used in the test. Results The positive rate of PCR test was 51.2% in 125 blood samples, and was significantly higher than the positive blood culture (25.6%), or the indicator of two abnormal non-specific parameters (32.8%) (P
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Objective To explore a rapid diagnostic method in neonatal sepsis and bacterial me- ningitis. Methods The primers and TaqMan probes were designed and synthesized based on the sequences of bacterial 16S rRNA gene. Nineteen bacterial strains, 3 different viruses, fungus and human genomic DNA were tested by FQ-PCR assay. Blood specimens and CSF from 195 cases of suspected septicemia were detected with both TaqMan PCR assay and blood or/and CSF culture. Results The FQ-PCR showed very high sensitivity and specificity and was able to detect at least 10 copies of 16S rRNA gene equivalent to 1~2 copies bacterium. No cross-reaction was found with human genomic DNA, other fungus and viruses. Among the 195 cases, the positive rate by FQ-PCR was 12.8%(25 cases) and 7.1%(15 cases) by blood culture ( P