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【Objective】 To investigate the diagnostic efficacy of a novel bladder cancer detection system utilizing a urine cell processing kit for urine sample preservation and detection. 【Methods】 Patients with primary persistent gross hematuria and high recurrence risk of bladder cancer after transurethral resection of bladder tumor were prospectively enrolled between Dec.2021 and Mar.2022. Urine specimens were either added to (experimental group) or not added to (control group) the urine cell processing kit and were fixed on Day 0, Day 3 and Day 7. The sensitivity and specificity of the two groups were compared after the cells were fixed, produced, stained and read with body fluid cytology total staining technique. 【Results】 The sensitivity and specificity of the experimental group on Day 0 were 82.50% (33/40) and 87.50% (14/16), respectively; those of the control group were 79.49% (31/39) and 82.35% (14/17), respectively. On Day 3, the sensitivity and specificity of the experimental group were 76.32% (29/38)and 81.25% (13/16), respectively; those of the control group were 52.78% (19/36) and 78.57% (11/14), respectively. On Day 7, the sensitivity and specificity of the experimental group were 71.43% (25/35) and 72.22% (13/18), respectively; those of the control group were 35.71% (10/28) and 60.00% (9/15), respectively. The sensitivity of the experimental group on Day 3 and Day 7 was significantly higher than that of the control group (P<0.05). 【Conclusion】 This bladder cancer urine cytology detection system provides clear diagnostic advantages and can be used as an auxiliary examination before cystoscopy for patients with hematuria and those at high risk of bladder cancer recurrence. It can also be used as a bladder cancer screening tool for pre-screening a large sample of people in order to achieve early diagnosis and treatment of bladder cancer.
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Objective To evaluated the HCV genotyping results which obtained by genotype diagnostic kit in Shenzhen area. Methods 158 samples which ELISA test of anti-HCV were positive were collected from voluntary blood donors from 2014 to 2015,and were tested by PCR fluorescence probe method for viral load.The samples which viral load were greater than 1.0 ×103 IU/mL were then tested by HCV RNA genotype diagnostic kit.To analysis the proportion of different genotypes and the correla-tion between genotypes with vrial load.Results 54 HCV RNA reactive sample were quantity by PCR fluorescence probe method from 158 anti-HCV positive samples.The genotyping data for 45 cases which vrial load greater than 1.0×103 IU/mL were obtained by HCV RNA genotype diagnostic kit.The frequencies HCV genotype 1b,2,3 and 6 were 57.78%(26/45),6.67%(3/45),8.89%(4/45)and 26.67%(12/45),respectively.One-way ANOVA analysis showed that significant difference in viral loads was found be-tween different HCV genotype 1b and 2(F =2.861,P <0.05),and there was a significant difference in viral loads and anti-HCV S/CO by sex(P <0.05).Fisher′s exact test showed the significance difference between age and genotypes(P <0.05 ).Conclusion HCV 1b and 6 were the most predominant genotypes due to the higher viral load than the other subtypes among volunteer blood do-nors in Shenzhen,while the proportion of HCV 2,3 declined.
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Objectives: To express and characterize NS1 of Indonesian-specific DENV2 virus in Pichia pastoris (P. pastoris). Methods: A codon optimized synthetic gene derived from the DENV-2 NS1 amino acid sequences was synthesized commercially and inserted into the P. pastoris pPICZαA expression vector. The recombinant DENV-2 NS1 protein was purified by Ni-NTA af-finity chromatography, and its antigenicity was tested. Results: The recombinant DENV-2 NS1 protein was secreted as a protein with a mo-lecular weight of ～45 kDa, and the optimal expression condition was achieved by in-duction with 2％(v/v) methanol for 72 h. The purified recombinant DENV-2 NS1 protein was able to interact with a monoclonal antibody of NS1 in a commercial rapid test. Conclusions: The resulting recombinant DENV-2 NS1 protein produced in P. pastoris KM71 is a potential candidate for use in the development of a dengue diagnostic kit and vaccine.
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Objectives To express and characterize NS1 of Indonesian-specific DENV2 virus in Pichia pastoris (P. pastoris). Methods A codon optimized synthetic gene derived from the DENV-2 NS1 amino acid sequences was synthesized commercially and inserted into the P. pastoris pPICZαA expression vector. The recombinant DENV-2 NS1 protein was purified by Ni-NTA affinity chromatography, and its antigenicity was tested. Results The recombinant DENV-2 NS1 protein was secreted as a protein with a molecular weight of ∼45 kDa, and the optimal expression condition was achieved by induction with 2% (v/v) methanol for 72 h. The purified recombinant DENV-2 NS1 protein was able to interact with a monoclonal antibody of NS1 in a commercial rapid test. Conclusions The resulting recombinant DENV-2 NS1 protein produced in P. pastoris KM71 is a potential candidate for use in the development of a dengue diagnostic kit and vaccine.
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Aims: To determine the effect of CD4 count (a glycoprotein found on the surface of immune cells such as T helper cells, monocytes, macrophages and dendritic cells) on the Candida species associated with Oropharyngeal candidiasis among HIV suspected patients. Place and Duration of the Study: State Hospital Ijebu Ode Ogun State Nigeria between February 2010 to August 2011. Methodology: Outpatients attending State Hospital Ijebu Ode were screened for HIV infection using Determine kits, Stat-pak kits and Unigold test kits. Western blot was used to confirm HIV infection and to determine the predominance of HIV specific glycoproteins in HIV seropositive patients. A total of 350 samples of sputum and blood from the HIV seropositive individuals while 300 samples from the HIV seronegative individuals. Sputum samples were cultured on sabouraud dextrose agar, and the isolates were Gram stained while the yeast-like fungi were subjected to germ tube test. CD4 count in blood samples was determined using flow cytometry. Results: HIV prevalence in females was 70.6% and in males was 29.4%. From three hundred and fifty patients suspected as HIV positive, seventy three had oral candidiasis (20.9%) while 277 (79.1%) were candidiasis negative. Higher oral candidiasis occurred in females (22.7%) than in males (16.5%). Candida alblicans was found to have higher occurrence of 86% among other Candida species. There is no significant association between the occurrence of oral candidiasis and the age of HIV subjects. There was higher occurrence of cases of immune depression (<350 CD4 count) in HIV seropositive (56.3%) than in HIV seronegative (0%) subjects. Candida infections occur when CD4 count was 200-500 cell/μl and usually represent the first indication of immune suppression. Decrease in CD4 count led to increase in occurrence of Candida species. The lowest number of Candida species was recorded when CD4 count was above 300 and Candida alblicans is the most predominant species isolated in this study. Conclusion: The result of this study shows that HIV infection led to decrease in CD4 count which in turn promotes candidiasis.
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Aim: To know whether one of the commercially available immunochromatographic tuberculosis tests is comparable with the widely available method, direct sputum microscopy. Design: The study prospectively validated the pulmonary tuberculosis rapid test kit using the reference standard, Lowenstein Jensen culture and compared the outcome with the direct sputum microscopy. Place and Duration: The study was conducted in Zankli Medical Centre, Abuja, between November 2004 and July 2005. Methodology: 340 patients from direct observation therapy clinics located in six different government owned health facilities were referred to our facility. These patients; male (192) and female (148) were between the age of 10 and 64 years old. Three sputa samples were collected over two consecutive days and direct microscopy and culture were performed on these samples. Also, 4ml of blood were collected from the same patients for antibody detection using immunochromatographic technique. Results: The evaluated rapid diagnostic kit when compared with the reference standard has a sensitivity of 59.3% and 81.1% specificity. Sensitivity and specificity of direct microscopy, when compared with the rapid test is statistically significant (P=0.001); indicating diagnostic accuracy of the conventional method of pulmonary tuberculosis testing over the immunochromatographic test. Conclusions: The conventional test indicated high performance in this report and it is suggestive of the relevance and diagnostic accuracy of the widely available method in the diagnosis of pulmonary tuberculosis in developing countries. This assertion is also, supported by the 2008 WHO/TDR report on evaluation of nineteen tuberculosis rapid diagnostic kits.
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This study aimed to produce immunoglobulin Y (IgY) specific to dengue virus which could be used for diagnostic kit of dengue. Lohman laying hens were immunized intramuscularly with antigenic of dengue. Egg yolk was separated from egg white and IgY antibody was then purified by multiple polyethylene glycol (PEG) 6000 extraction and ammonium sulfate purification steps. The IgY concentration in egg yolk increased during the immunization period until week 6 where it began to increase dramatically at 2 weeks and it reached a plateau at 4 weeks after immunization. After week 6 the levels decreased gradually. Antibody of dengue was detected and produce a specific line of precipitation in agar gel precipitation test (AGPT) beginning the second week after the first immunization. Analysis of results obtained with ELISA showed significant increase in the denguespecific antibodies after two weeks from the primary immunization. Through the effect of boostering; the antidengue antibody levels reached a plateau at four weeks from the primary immunization and remained significantly higher till the end of observation period. SDS-PAGE revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 145; 66; 45, 33 and 26 kDa and western blot analysis revealed the presence of anti-dengue IgY in egg yolks protein, with a molecular weights of approximately 66 kDa. These results suggested that chicken IgY could be a practical strategy in large-scale production of specific anti-dengue Ig Y for diagnostic KIT of dengue.
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Background: IgM antibody capture ELISA (MAC-ELISA) technique has been widely applied for Japanese Encephalitis Virus (JEV) diagnosis. So far rare internationally commercial kits are available. Thus, the international evaluation of the kit is required as per the recommendation of the WHO. Objectives: To evaluate the quality of the IgM antibody capture ELISA diagnostic kit for JEV produced by the Vietnam National Institute of Hygiene and Epidemiology (NIHE). Subjects and method: In this study, NIID kit was used as control to check the kit from NIHE. Both NIHE and NIID kits were used to detect JEV IgM among 38 serum and 6 CFS samples, which belongs to 5 sample groups (JE patients group, dengue patients group, other viral encephalitis patients group, Tick Born Encephalitis (TBE) patient group and healthy JE vaccinated donors group). Results: The detection of JEV IgM by NIHE kit was concurrent with the NIID kit. There is no positive with the JE in the groups of Dengue patients, TBE, other virus encephalitis patients and JE vaccinated donors. Conclusion: MAC-ELISA kit of NIHE can be used for different diagnosis of JEV and Dengue virus (both viruses are in Flavivirus genus), as well as other viruses caused by encephalitis.
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Inmunoglobulina M , Virus de la Encefalitis Japonesa (Especie)RESUMEN
Objective:Use a new F-PCR method to develop a hepatitis C virus(HCV) diagnostic kit, test the kit through clinical trial and compare it with immunological method. Methods: Fluorescence PCR(F-PCR) is a method which combines PCR and fluorescence probe hybridization together to measure DNA/RNA. Because in-tube monitoring of fluorescence signal can be done to stand for the quantitity of PCR product. Electrophoresis and UV detection are eliminated, so after-PCR cross-contamination which causes false positive can be avoided. Results:A clinical diagnostic kit for HCV with this method is developed. 512 clinical serum samples were tested with this kit, using HCV FLISA kit from Abbott Co.and HCV Fluorescence RT-PCR kit from Biotronics Co. (B-PCR) as control. The results shows :positive rate is 30.5%,sensitivity 97.3 % and specificity 98.1% . Conclusion: F-PCR is obviously superior to ELBA, and higher than B-PCR in sensitivity. The specificity of those three kits have no statistic difference. F-PCR can be used to monitor RNA of HCV in serum, and could be useful for clinical diagnose and therapy effects monitoring.