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.

Objective:To study the effect of endogenous IL-1? on the expression of matrix metalloproteinases (MMPs) and COX2 in dental pulp cells. Methods:Human dental pulp cells were treated with human recombinant IL-1? at 1 nmol/L in serum-free medium for 18 h. Then the cells were collected and total RNA was isolated, MMPs and COX2 mRNA expression was assessed by semiquantitative RT-PCR. Results:IL-1? at 1 nmol/L induced the expression of COX2 and MMP-1 mRNA in human dental pulp cells. Conclusion:IL-1? may contribute to stimulating expression of MMPs and COX2 in the dental pulp during pulpitis.

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.

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.

Objective To observe the effects of 3,4,5-trihydroxybenzoic acid dimmer (TAD) in different doses on apoptosis and cell cycle progression of liver cancer SMMC-7721 cells in vitro and explore its possible mechanism of inhibiting tumor growth.Methods The experiment was divided into 0,3.125,6.250,12.500 and 25.000 ?g TAD groups. Apoptosis was observed with JEM-1200EX transmission electron microscope (TEM).The changes of apoptosis and cell cycle progression were measured with flow cytometry.Results The typical features of apoptosis were found in tumor cells,the nuclei were broken to pieces,mitochondrion cristae were disrupted and vacuoles were showed in nucleus and cytoplasm in 25.000 ?g TAD group observed under TEM.Meantime,the apoptotic percentages of the cells treated with 3.125—25.000 ?g TAD were increased significantly as compared with control group (P

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.

Objective To analyze the antigens of solid cultured germ, liquid cultured germ and secreted composition, looking for the main immunity antigens of Mycobacterium tuberculosis H37Rv.MethodsShowing the difference of the antigens of solid cultured germ, liquid cultured germ and secreted composition by SDS-PAGE, Western blotting, monoclonal and polyclonal antibody techniques.Results Antigens of the solid cultured germ were nearly accordance with the liquid cultured germ and they were difference with secreted proteins. The main immunity antigens of the cultured germ were 79 000, 54 000 ,38 000~50 000, 28 000 and the secreted proteins are 66 000, 55 000~64 000, 30 000~34 000, 24 000, 16 000.The antigens of 66 000, 55 000 and 16 000 were proved that they were secreted proteins by monoclonal antibodies.Conclusions This study demonstrated that the main immunity antigens of the cultured germ in Mycobacterium tuberculosis H37Rv were difference with the secreted one .This is beneficial to the cognition of H37Rv and looking for special antigens for diagnosis of tuberculosis .

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.

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.

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.