Evaluation of Interferon-Gamma Release Assay Testing and Tuberculin Skin Test for Early Diagnosis of Tuberculosis in Children and Adolescents

Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country with a high burden of tuberculosis. Method: This study included 581 children and adolescents aged 4 - 19 years who were suspected of having tuberculosis, were latently infected with Mycobacterium tuberculosis, and had received at least one dose of BCG vaccine between April 17, 2019, and February 24, 2021. The study evaluated the TST results of 106 patients who had a positive Quantiferon test and were suspected of having tuberculosis. Results: The study included 581 patients aged between 4 and 19 years. Of these, 106 patients tested positive for the Quantiferon test, while 19 were indeterminate and 456 were negative. The Quantiferon test positivity rate was 18.24%. Among the 106 QFT-Plus-positive cases, 23 patients also tested positive for TST. The difference in distribution was found to be statistically significant. Conclusion: The QFT-Plus test is considered an alternative to TST and other microbiological diagnostic methods for early tuberculosis diagnosis, particularly in children and adolescents.

Study of Low-intensity 2450-MHz Microwave Exposure Enhancing the Genotoxic Effects of Mitomycin C Using Micronucleus Test and Comet Assay in vitro

Objective To determine the interaction between 2450-MHz microwaves (MW) radiation and mitomycin C (MMC). Methods The synergistic genotoxic effects of low-intensity 2450-MHz microwave and MMC on human lymphocytes were studied using single cell gel electrophoresis (SCGE) assay (comet assay) and cytokinesis-blocked micronucleus (CBMN) test in vitro. The whole blood cells from a male donor and a female donor were either only exposed to 2450-MHz microwaves (5.0 mW/cm2) for 2 h or only exposed to MMC (0.0125 μ/mL, 0.025 μg/mL, 0.05μg/mL and 0.1 μg/mL) for 24 h; and the samples were exposed to MMC for 24 h after exposure to MW for 2 h. Results In the comet assay, the comet lengths ( 29.1 μm and 25.9 μm) of MW were not significantly longer than those (26.3 μrn and 24.1 μm) of controls (P>0.05). The comet lengths (57.4 μm, 68.9 μm, 91.4 μm, 150.6μm and 50.6 μm, 71.7μm, 100.1 μm, 145.1 μm) of 4 MMC groups were significantly longer than those of controls (P<0.01). The comet lengths (59.1 μm, 92.3 μm, 124.5 μm, 182.7 μm and 57.4 μm, 85.5 μm, 137.5 μm, 178.3 μm) of 4 MW plus MMC groups were significantly longer than those of controls too (P<0.01). The comet lengths of MW plus MMC groups were significantly longer than those of the corresponding MMC doses (P<0.05 or P<0.01) when the doses of MMC were ≥50.025 μg/mL. In the CBMN, the micronucleated cell (MNC) rates of MW were 5% and 6%, which showed no difference compared with those (4‰ and 4‰) of controls (P>0.05). The MNC rates of 4 MMC groups were 8‰, 9‰, 14‰, 23‰ and 8‰, 8‰, 16‰, 30‰ respectively. When the doses of MMC were 3≥0.05 μg/mL, MNC rates of MMC were higher than those of controls (P<0.05). MNC rates of 4 MW plus MMC groups were 12‰, 13‰, 20‰, 32‰ and 8‰, 9‰, 23‰, 40‰. When the doses of MMC were 5≥0.05 μg/mL, MNC rates of MW plus MMC groups were much higher than those of controls (P<0.01). MNC rates of 4 MW plus MMC groups were not significantly higher than those of the corresponding MMC doses. Conclusion The low-intensity 2450-MHz microwave radiation can not induce DNA and chromosome damage, but can increase DNA damage effect induced by MMC in comet assay.

Joannesia princeps： Evaluation of Aqueous Extracts Genotoxicity Utilizing Allium cepa Assay and Micronucleus Test

Evaluation of the genotoxic and cytotoxic potentials is a part of the analysis process where plant products with therapeutic properties must be submitted for being safely employed, mainly through long time points, doannesia princeps, also known as ＂cotieira＂ has been used on popular medicine as laxative and several diseases treatment. Aiming to analyze genotoxic potential ofd. princeps leaves extract, Allium cepa assay and micronucleus test were employed. No genotoxic activity presented J. princeps leaves extract; however, extracts cytotoxic activity over A. cepa meristematic cells and mice marrow cells was observed. J. princeps leaves extract presented antiproliferative activity at concentrations and systems employed indicating their therapeutic potential for cellular cycle inhibition. Moreover, results showed that in regard to its mutagenic effect, usage of J. princeps leaves tea on popular medicine has been a safe procedure, as well.

Real-world utility of serological tests in patients with suspected scrub typhus in the Republic of Korea:A single-center,retrospective,observational study

Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult patients with suspected scrub typhus who visited a tertiary care hospital in the Republic of Korea from September to December from 2019 to 2021.The included patients had an acute fever and at least one of the following ten secondary findings:myalgia,skin rash,eschar,headache,thrombocytopenia,increased liver enzyme levels,lymphadenopathy,hepatomegaly,splenomegaly,and pleural effusion.The diagnoses were grouped as scrub typhus or other diseases by two infectious disease physicians.Results:Among 136 patients who met the eligibility criteria,109 had scrub typhus and 27 had different diseases.Single and paired total antibodies using immunofluorescence assay(IFA),and total antibodies using immunochromatography-based rapid diagnostic testing(ICT)were measured in 98%,22%,and 75%of all patients,respectively.Confirmation using paired samples for scrub typhus was established at a median of 11[interquartile range(IQR)10-16]days following the first visit.Among the 82 admitted patients,the median admission time was 9(IQR 7-13)days.According to IFA,58(55%)patients with scrub typhus had total immunoglobulin titers≥1:320,while 23(85%)patients with other disease had titers<1:320.Positive ICT results were observed in 64(74%)patients with scrub typhus and 10(67%)patients with other diseases showed negative ICT results.Conclusions:Serological testing for scrub typhus is currently insufficient for decision-making in clinical practice.

The Cytokinesis-Block Micronucleus Assay on Peripheral Blood Lymphocytes as a Prospective Biological Test-System to Estimate the Influence of Radon on the Human Organism: Recent Progress and Future Prospects

This paper discusses the problem of assessing the negative after-effects of low doses of ionising radiation exposure in humans. Radon and its decay daughter products are the most widespread source of such irradiation. Miners (in both uranium and non-uranium mines) as well as laypeople in domestic life may be exposed to radon, making the problem of assessing the cytogenetic effects of exposure extremely crucial. One of the more promising test systems to assess the effect of radon is the cytokinesis-block micronucleus assay (CBMN) on peripheral blood lymphocytes, which has a number of advantages over other cytogenetic techniques. Recent progress and future prospects of this cytogenetic method are discussed here.

Predictive value of MTT assay as an in vitro chemosensitivity testing for gastric cancer:One institution's experience

AIM:To investigate the predictive clinical value of in vitro 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay for directing chemosensitivity in patients with gastric cancer. METHODS:Results of a total of 353 consecutive patients with gastric cancer treated with MTT-directed chemotherapy or physician’s empirical chemotherapy from July 1997 to April 2003 were reviewed and analyzed retrospectively. RESULTS:The overall 5-year survival rate of MTT- sensitive group (MSG) and control group (CG) was 47.5% and 45.1%, respectively. The results of subgroup analysis with Cox proportional-hazards model were favorable for the MSG-sensitive group. However, no statistically significant difference in survival rate was observed between the two groups. CONCLUSION:Individualized chemotherapy based on in vitro MTT assay is beneficial, but needs to be confirmed by further randomized controlled trials.

Detection of Cytogenetic Effects in Peripheral Lymphocytes of Students Exposed to Formaldehyde with Cytokinesis-Blocked Micronucleus Assay

Cytokinesis-blocked micronucleus assay was applied as a biological dosimeter to detect abnormalities in human peripheral lymphocytes of thirteen students exposed to formaldehyde (FA) during a 12-week (10 h per week) anatomy class. Breathing-zone air samples colleeted during dissection procedures showed a mean concentration of 2. 37 ppm (3. 17mg/m3 ). Ten students from the same school but without FA exposure served as controls. Chromosome aberrations (CA) and sister chromatid exchanges (SCE) were detected in both groups. The micronuclei (MN) rate (6. 38 ± 2. 50‰ ) and CA rate (5. 92 ±2. 40‰ ) in the FA-exposed group showed a significant increase (P< 0. 01 ) when compared with those of the controls (3. 15 ±1. 46‰and 3. 40 ± 1. 57 % respectively). A correlation between MN and CA in individuals was observed. SCE in the exmpd group were also increased (P< 0. 05), but not so greatly as MN or CA. The results indicated that FA might damage the chromosomes of human lymphocytes.

Comparison of a rapid diagnostic test and microimmunofluorescence assay for detecting antibody to Orientia tsutsugamushi in scrub typhus patients in China

Objective:To evaluate the detection of IgM and IgG antibodies to Orientia tsutsugamushi(O. tsutsugamushi) by rapid diagnostic test(RDT) and microimmunofluorescence assay(ml FA). Methods:RDT using a mixture of recombinant 56-kDa proteins of O.tsutsugamushi and mIFA assay were performed on 20 patients from Fujian and 13 patients from Yunnan Province,and 82 sera samples from healthy farmers in Anhui Province and Beijing City in 2009.Comparison of the RDT and mIFA assay was performed by using X test and the P level of 【0.05 was considered to be significance.Results:Among these 82 normal sera samples,the specificity of RDT was 100%for both IgM and IgG tests.In 33 samples from patients with scrub typhus,5 cases were positively detected earlier by RDT than by mIFA in IgM test,and 2 cases were positive in IgG test.Sensitivities of RDT were 93.9%and 90.9%for IgM and IgG,respectively.The sensitivity of combination lest of IgM and IgG was 100%.Geometric mean titer diluted sera from confirmed cases by IFA and RDT assay were 1:37 vs.1:113(P【0.001) in IgM test and 1:99 vs.1:279 (P【0.05) in IgG test.Conclusions:RDT is more sensitivite than mIFA in the early diagnosis of scrub typhus and it is particularly applicable in rural areas.

NASOPHARYNGEAL CARCINOMA RADIOSENSITIVITY PREDICTION BY CYTOKINESIS－BLOCK MICRONUCLEUS ASSAY

Cytokinesis-block micronucleus method is used to evaluate the radiosensitivity of a nasopharyngeal carcinoma cell line (CNE-1) and biopsies obtained from 31 patients with nasopharyngeal carcinoma. The number of micronuclei increases with theradiation dose. A good correlation was found between the radiosensitivity determined by the micro-nucleus assay and that measured by the colony-forming assay in CNE-1 cell line (r=-0.998). Moreover, the results of micronucleus assay for tumor cells from biopsies of patients with primary carcinoma look promising for the prediction of tumor radiosensitivity. These results are encouraging but fleed to be confirmed with a larger number of patients with a longer follow-up.

The Chromosomal Effect of Birchen Dust as Determined by the Micronucleus Test

In a wood processing factory, the measured air concentration of birchen dust was 1. 26 ±0. 41 mg/m3, and the micronucleus frequency of peripheral blood lymphocytes in 83 workersexposed to wood dust was 1. 13 ± 2. 83%, which was significantly higher (P < 0.01 ) thanthat of control group (0. 51 ± 1. 41% ). The number of exposed workers with positive mi-cronucleus test was 9. 6 %, which was higher than that of control group (4. 5 % ), but thedifference was not significant (P >0. 05 ). The micronucleus test in mice treated with waterextracts of unsteamed and unbaked birchen dust showed that the micronucleus frequencies inall treated groups were significantly higher than that of contro group (P < 0. 01 ) and therewas also a doseresponse correlation (r = 0. 96, P < 0. 0005 ). The results of steamed andbaked birchen dust extracts were significantly lower than those of the unsteamed and unbakedones at the same doses (P< 0. 001 ). This suggests that when the birchen dust is steamed atthe temperature of 100℃ for 24h or baked at the temperature of 80℃, its inducing effect inmicronucleus test could be lowered

Evaluation of Protective Effects of Bioactive Phytochemicals Against Methotrexate in Salmonella typhimurium TA1535/pSK1002 Coupled with Micronucleus Assay

We evaluated the antimutagenic effects kinds of bioactive phytochemicals and of 10 some phytochemical combinations against methotrexate （MTX）-induced genotoxicity by the umu test in Salmonella typhimurium TA1535/rpSK1002 combined with a micronucleus assay. We observed that allicin, proanthocyanidins, polyphenols, eleutherosides, and isoflavones had higher antimutagenic activities than the other five types of bioactive phytochemicals. At the highest dose tested, MTX-induced genotoxicity was inhibited by 25%-75%. Kunming mice treated by MTX along with bioactive phytochemical combinations showed significant reduction in micronucleus induction and sperm abnormality rate （P〈0.01）. These results indicate that bioactive phytochemical combinations can be potentially used as new cytoprotectors.

CUMULATIVE RESULTS OF CHEMOSENSITIVITY TEST USING MTT ASSAY IN DOUBLE-LAYER AGAROSE

Objective To investigate cumulative results of chemosensitivity test using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl （MTT） assay in double-layer agarose. Methods A total of 2 491 patients with different kinds of cancers were enrolled in the study, in which 18 kinds of different anticancer drugs were used. A computer soft was used to get charts. ResultsThe total evaluability rate was 82.7% （2 060/2 491）. Among all agents, the efficiency rates of 5-Fu, MMC, DDP, BLM and CBP were higher than the efficiency rates of others. The response rate range of different cancer in vitro sensitivity by using MTT assay in double layer agarose were from 9.2% （biliary duct） to 37.5% （malignant lymphoma）. For colon and rectum cancer, 5-Fu, DDP, MMC and BLM were more sensitive than other anti-tumor agents. For breast carcinomas, ACTD and DDP were more sensitive. For gastric cancer, 5-Fu, DDP and BLM were more sensitive. For leukemia, VM-26 and HHRT were more sensitive. ACM was more sensitive to kidney and MXT and BLM were more sensitive to pancreas cancer. For Lung cancer, DDP and EPI were more sensitive. Mean true positive rate, mean true negative rate, mean sensitivity, mean specificity and mean accuracy were 44% , 92% , 72% , 77% , and 76% , respectively. Conclusion Chemosensitivity tesing using the MTT assay in a double layer agarose was a very useful reference to chem- therapy.

基于4%中性甲醛固定肝组织的大鼠肝微核实验方法的建立与验证

目的建立并验证基于4%中性甲醛固定肝组织制备肝细胞的大鼠肝微核实验(LMNT)方法(甲醛固定-LMNT法)。方法(1)SD大鼠分为雌性和雄性组,然后分别随机分为溶剂对照组和肝微核阳性化合物N-亚硝基二乙胺(DEN)12.5 mg·kg^(-1)组,每组5只,分别ig给予生理盐水和DEN,每天1次,连续14 d后取肝组织,同时用胶原酶消化-LMNT法和甲醛固定-LMNT法检测微核化肝细胞数量和处于有丝分裂期肝细胞数,分别计算肝细胞微核率和有丝分裂指数,肝细胞微核率>0.07%判定为阳性结果。(2)雄性SD大鼠分为喹啉(30,60和120 mg·kg^(-1))组、N-亚硝基吡咯烷(NPYR,25,50和100 mg·kg^(-1))组、各自溶剂对照组(NPYR,去离子水;喹啉,玉米油)及阳性对照DEN(12.5 mg·kg^(-1))组,每组5~6只,连续ig 15 d。每日记录体重,实验结束取肝记录肝总重,计算肝系数。自动生化分析仪检测肝功能相关血清生化指标谷丙转氨酶(GPT)、谷草转氨酶(GOT)活性和血清总胆红素(T-BIL)、直接胆红素(D-BIL)的水平。采用胶原酶消化-LMNT法和甲醛固定-LMNT法测定肝细胞微核率,用外周血微核实验和肝彗星实验评价喹啉和NPYR遗传毒性。结果(1)与各自溶剂对照组的肝细胞微核率(0.069%和0.030%)相比,甲醛固定-LMNT法测得的DEN组雄性大鼠肝细胞微核率为1.10%,雌性大鼠为0.82%,均显著升高(P<0.05);与各自溶剂对照组(0.060%和0.030%)相比,胶原酶消化法-LMNT法测得的DEN组雄性大鼠肝细胞微核率为1.45%,雌性大鼠为0.46%,均显著升高(P<0.05),判定为阳性。2种方法中雄性大鼠的肝细胞微核率均显著高于雌性(P<0.05)。与各自溶剂对照组相比,2种方法测得的DEN组雄性和雌性大鼠肝细胞有丝分裂指数均无明显差异。(2)与溶剂对照组相比,NPYR 50和100 mg·kg^(-1)组大鼠体重在ig第7~14天显著降低(P<0.01),DEN组在ig第8~14天显著降低(P<0.01);喹啉120 mg·kg^(-1)组大鼠在ig第4~14天显著降低(P<0.01),DEN组在ig第10~14天显著降低(P<0.05)。与溶剂对照组相比,NPYR 100 mg·kg^(-1)组(P<0.01)和DEN组(P<0.05)的肝重和肝系数均显著降低;喹啉60和120 mg·kg^(-1)组肝重(P<0.01)和肝系数(P<0.05)均显著增加。与溶剂对照组相比,DEN组大鼠血清T-BIL水平显著升高(P<0.01),NPYR 100 mg·kg^(-1)组GPT、GOT活性和D-BIL、T-BIL水平均显著升高(P<0.01),NPYR 25,50 mg·kg^(-1)和喹啉各剂量组则均无显著差异。甲醛固定-LMNT法测得的NPYR组肝细胞微核率较胶原酶消化-LMNT法略高,均判定为阳性;与溶剂对照组相比,2种方法测得的NPYR组肝细胞微核率均显著增加(P<0.05),喹啉组结果相当,均判定为阳性;2种方法测得的喹啉组肝细胞微核率均显著增加(P<0.05)。2种方法测得的NPYR和喹啉组肝细胞微核率相关性较好(R2分别为0.8614和0.9279)。NPYR组外周血微核实验为阴性,彗星实验结果为阳性;喹啉组外周血微核实验和彗星实验结果均为阴性。结论初步建立并验证了甲醛固定-LMNT法,且该方法检测肝致癌物的灵敏度与胶原酶-LMNT法相近。

血栓通注射液临床前遗传毒性研究

目的 采用细菌回复突变试验(Ames试验)、CHL细胞染色体畸变实验和小鼠骨髓细胞微核试验,综合评价血栓通注射液的遗传毒性。方法 Ames试验,在有或无S9活化条件下,血栓通注射液设置128.0、320.0、800.0、2000.0、5000.0 mg每皿共5个浓度组,用平皿掺入法检测菌株TA1535、TA97、TA98、TA100和TA102的致突变性。CHL细胞染色体畸变试验,血栓通注射液设置125、250、500μg·m L^(-1)共3个浓度组,分别对CHL细胞染毒4 h(在有或无S9活化条件下)和24 h(在无S9活化条件下)。油镜下观察300个中期分裂相细胞中含结构畸变和数目畸变的细胞数。小鼠骨髓细胞微核试验,血栓通注射液设置104.3、208.6、417.2 mg·kg^(-1)共3个剂量组。KM小鼠经尾静脉注射给药2 d,末次给药后24 h取材,计数嗜多染红细胞(PCE)中的微核数和PCE比例。结果 血栓通注射液各处理组测试菌株回复突变菌落数与溶媒对照组相比均未见增加1倍以上。血栓通注射液各处理组的结构畸变率和数目畸变率与溶媒对照组比较均无明显差异(P>0.05)。各剂量组小鼠骨髓细胞平均含微核嗜多染红细胞率与溶媒对照组比较均无明显差异(P>0.05)。结论 本实验室确定的条件下,血栓通注射液的遗传毒性评价结果为阴性。

两种染色体收获方法在淋巴细胞微核试验中的价值比较

目的:探讨改良后淋巴细胞收获法及滴片方法在淋巴细胞微核试验中的优势。方法:分别用改良后淋巴细胞收获及滴片方法(试验组)和常规法(对照组)收获淋巴细胞,经滴片、染片并统计分析两种方法各种淋巴细胞的微核的检出率及工作效率。结果:试验组用时(180min)小于对照组(320min),试验组得到的微核率为(2.23‰±2.74‰)、微核细胞率为(1.6‰±1.96‰)和微核阳性率为(57.5‰)均高于对照组(1.57‰±2.3‰,1.25‰±1.73‰,46.6%),差异均有统计学意义(P<0.01,P<0.01,P<0.05)。结论:改良法(试验组)明显缩短了完成试验所用的时间,简化操作步骤,同时减少了收获过程中对细胞的机械损伤,使淋巴细胞微核的检出率得到有效提高,为临床医生提供了更为可靠的诊断依据。

Effect of various drinking water on human micronucleus frequency in high-risk population of PHC

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Study on Colloidal Gold Immunochromatography Assay for Rapid Detection of Spectinomycin

[Objectives] This study was conducted to establish a rapid detection method for spectinomycin in pork,chicken,fish,shrimp flesh and water.[Methods]A test strip for rapid detection of spectinomycin in milk was developed by colloidal gold immunochromatography assay. [Results]The test strip had a detection limit of 50 μg/kg to milk with a detection time of 15 min,and the false positive rate and false negative rate were both 0. [Conclusions]The method is accurate,simple,reliable and convenient,and is suitable for rapid on-site spectinomycin detection.

Response of Lymphocytes to Radiation in Untreated Breast Cancer Patients as Detected with Three Different Genetic Assays

Objective To detect the response of lymphocytes to radiation in untreated breast cancer patients with three different genetic assays. Methods Blood samples were collected from 25 untreated patients and 25 controls. Each blood sample was divided into two parts： one was irradiated by 3-Gy X-ray （irradiated sample）, the other was not irradiated （non-irradiated sample）. The radiosensitivity of lymphocytes was assessed by comet assay, cytokinesis-block micronucleus （CBMN） assay and 6-TG-resistant cells scored （TG） assay. Results The baseline values of micronucleated cell frequency （MCF） and micronucleus frequency （MNF） in the patients were significantly higher than those in the controls （P〈0.01）, and 3-Gy X-ray induced genetic damage to lymphocytes in the patients increased significantly as compared with that in the controls as detected with the three genetic assays （P〈0.01）. The proportion of radiosensitive cases in the patient group was 48% for the mean tail length （MTL）, 40% for the mean tall moment （MTM）, 40% for MCF, 44% for MNF, and 48% for mutation frequencies of the hprt gene （Mfs-hprt）, respectively, whereas the proportion of radiosensitive cases in the control group was only 8% for all the parameters. Conclusion The difference in the lymphocyte radiosensitivity between the breast cancer patients and the controls is significant. Moreover, there are wide individual variations in lymphocyte radiosensitivity of patients with breast cancer. In some cases, the radiosensitivity of the same patient may be different as detected with the different assays. It is suggested that multiple assays should be used to assess the radiosensitivity of patients with breast cancer before therapy.

Hepatitis C virus antigens enzyme immunoassay for one-step diagnosis of hepatitis C virus coinfection in human immunodeficiency virus infected individuals

BACKGROUND Current diagnosis of hepatitis C virus(HCV)infection requires two sequential steps:testing for anti-HCV followed by HCV RNA PCR to confirm viremia.We have developed a highly sensitive and specific HCV-antigens enzyme immunoassay(HCV-Ags EIA)for one-step diagnosis of viremic HCV infection.AIM To assess the clinical application of the HCV-Ags EIA in one-step diagnosis of viremic HCV infection in human immunodeficiency virus(HIV)-coinfected individuals.METHODS The study blindly tested HCV-Ags EIA for its performance in one-step diagnosing viremic HCV infection in 147 sera:10 without HCV or HIV infection;54 with viremic HCV monoinfection;38 with viremic HCV/HIV coinfection;and 45 with viremic HCV and non-viremic HIV coinfection.RESULTS Upon decoding,it was 100%accordance of HCV-Ags EIA to HCV infection status by HCV RNA PCR test.In five sera with HCV infection,HCV RNA was as low as 50-59 IU/mL,and four out of five tested positive for HCV-Ags EIA.Likewise,it was also 100%accordance of HCV-Ags EIA to HCV infection status by HCV RNA PCR in 83 sera with HCV and HIV coinfection,regardless if HIV infection was active or not.CONCLUSION The modified HCV-Ags EIA has a lower detection limit equivalent to serum HCV RNA levels of approximately 100 IU/mL.It is highly sensitive and specific in the setting of HIV coinfection,regardless of HIV infection status and CD4 count.These data support the clinical application of the HCV-Ags EIA in one-step diagnosis of HCV infection in HIV-infected individuals.

Fast and Accurate Identification of <i>M. tuberculosis</i>Complex Using an Immunochromatographic MPT64 Antigen Detection Test

Background: A new rapid Immunochromatographic test (ICT) kit (MPT64 TB Ag Kit) for detection of MPT64 Antigen in M. tuberculosis (MTB) isolates used for rapid identification of MTB isolates developed by SD (Standard Diagnostics) Bio line, South Korea was evaluated. The ICT is a rapid, reliable and cheaper method that can be used instead of conventional biochemical tests for confirming MTB in culture isolates in resource limited laboratories. The study also evaluated the ability of ICT to detect MPT64-Antigen before the micro MGIT could signal positive. Material/Methods: A total of 450 sputum samples of individual patients were used for the study. 152 isolates of Mycobacteria were recovered from solid and liquid media. These strains were tested for the detection of MPT64-antigen. H37Rv strain was served as the positive reference control and also used for early detection of Antigen experiment. Findings: The development of bands on both test and sample region when H37Rv strain was tested were seen (MPT64 antigen positive). When 138 MTB isolates were tested, it showed a similar banding pattern indicating 100% sensitivity. MPT64 band formation was not detected in any of the 14 isolates indicating 100% specificity. Both PPV & NPV were 100%. All the isolates negative for MPT64 Ag were confirmed as MOTT by conventional bio-chemical PNBA. The H37Rv strain showed a faint band from the 2nd day onwards from inoculation till 3rd day in the earlier Antigen detection experiment. Conclusion: Rapid identification of MTB culture isolate is a pressing need for diagnosis and proceeding to perform drug susceptibility testing. MPT64 TB Ag detection ICT kit is a rapid, reliable method, good substitute for molecular identification methods, and conventional biochemical test which is time-consuming and technically demanding. The early detection of Antigen can be used as an effective tool in diagnosis.