Displaying publications 41 - 60 of 127 in total

Abstract:
Sort:
  1. Fulltext The role of blood transfusion in HCV infection: a study testing Ab: RNA & genotype
    Al-Kubaisy, Waqar A., Niazi, Amjad D.
    Int J Public Health Res, 2011;1(2):72-78.
    MyJurnal
    Introduction Hepatitis C Virus (HCV) recently was identified as a major cause of post transfusion hepatitis world wide. To evaluate the role of blood transfusion on the prevalence of HCV infection, by testing antibody and RNA as well as the genotypes of HCV .Also to detect if Blood transfusion acts as unconfounding risk factor for HCV infection.
    Methods Sera from 3491 pregnant women were investigated for the presence of HCV antibodies (anti-HCV) by using third generation enzyme immunoassay (EIA-3) as screening test, followed by immunoblot assay (Lia Tek-III). In addition 94 sera of studied women were subjected to molecular analysis (at laboratories of Sorin BioMedica - Italy) for the detection of viral RNA and genotypes of HCV. Using RT-PCR & DNA Enzyme immunoassay (DEIA) method.
    Results Our study revealed, that seroprevalence rate of HCV specific Ab & RNA were significantly higher (16.32 %, 80% respectively) among women with a history of blood transfusion, compared to those (2.53%, 56.5%) with no such history P=0.0001, P=0.01. And there is a significant direct linear correlation between number of blood transfused and the seropositive rate of anti-HCV (r=0.7, p=0.046). Based on multivariate analysis, interestingly, this study confirmed that, blood transfusion significantly acting as unconfounding risk factor for acquiring HCV infection (Adjusted OR=1.938,95% C.I=1.646-2.28). And the risk of exposure is increases with increased number of blood transfused. Although, we found no significant association between, HCV genotypic distribution and history of blood transfusion. However, high proportion of women with a history of blood transfusion were harboring HCV genotype -4 or 1b, 50%,40%, resepctively.
    Conclusions Our study shows, evidence that, blood transfusion acts as unconfounding risk factor for acquiring and in a mode of transmission of HCV infection. Therefore strict screening of blood donor for HCV-Abs and / or RNA is highly recommended.
    Matched MeSH terms: Immunoenzyme Techniques
  2. The expression of Bcl-2 by proliferating cells varies in different categories of B-cell lymphoma
    Masir N, Jones M, Lee AM, Goff LK, Clear AJ, Lister A, et al.
    Histopathology, 2010 Apr;56(5):617-26.
    PMID: 20459572 DOI: 10.1111/j.1365-2559.2010.03524.x
    To investigate the relationship between Bcl-2 protein expression and cell proliferation at single-cell level in B-cell lymphomas using double-labelling techniques.
    Matched MeSH terms: Immunoenzyme Techniques
  3. Fulltext Detection of Toxoplasma gondii DNA by PCR following microwave treatment of serum and whole blood
    Meganathan P, Singh S, Ling LY, Singh J, Subrayan V, Nissapatorn V
    Southeast Asian J. Trop. Med. Public Health, 2010 Mar;41(2):265-73.
    PMID: 20578507
    Detection of Toxoplasma gondii in blood by means of the polymerase chain reaction (PCR) may facilitate early diagnosis of toxoplasmosis in different groups of patients. We evaluated this approach in 42 patients presenting with ocular or psychotic diseases by comparing the sensitivity and specificity of PCR after heat treatment using a microwave oven with a standard genomic DNA extraction method for paired serum and whole blood samples. The presence of serum IgM and IgG antibodies against T. gondii was detected using a standard commercial enzyme-linked immunosorbent assay and enzyme immunoassay for IgG avidity test. Of 42 whole blood samples, PCR after microwave treatment was positive in 8 samples with a sensitivity of 73% and specificity of 100% compared to 11 samples positive by the extraction method. Although none of 42 sera samples was PCR positive by the extraction method, 7 specimens were positive after microwave treatment. This is the first study to use a microwave heat treatment, which is simple, rapid and a promising alternative method, in detecting small amounts of T. gondii DNA in human blood. Furthermore, irradiation of blood samples with microwaves allows incorporation of PCR into a practical tool for routine clinical assessment of patients with Toxoplasma infection.
    Matched MeSH terms: Immunoenzyme Techniques
  4. Expression and alteration of p16 in diffuse large B cell lymphoma
    Lee ES, Kim LH, Abdullah WA, Peh SC
    Pathobiology, 2010;77(2):96-105.
    PMID: 20332669 DOI: 10.1159/000278291
    This study aimed to examine (1) the expression of P16 protein relative to sites of presentation, immunophenotypic subgroups and proliferative indices of tumour cells, and (2) the relationship between p16 gene alterations and P16 protein overexpression in 70 cases of diffuse large B cell lymphoma (DLBCL).
    Matched MeSH terms: Immunoenzyme Techniques
  5. Fulltext Role of fibrinolytic markers in acute stroke
    Abdullah WZ, Idris SZ, Bashkar S, Hassan R
    Singapore Med J, 2009 Jun;50(6):604-9.
    PMID: 19551314
    The fibrinolytic system plays an important role in normal haemostasis and endothelial function. This study was conducted to compare three fibrinolytic markers, i.e. plasminogen, tissue-plasminogen activator (t-PA) and plasminogen activator inhibitor type-1 (PAI-1) between acute stroke and stable non-stroke patients and to investigate the clinical significance of these markers.
    Matched MeSH terms: Immunoenzyme Techniques
  6. Comparison of histopathological features of Vibrio cholerae O1 El Tor and O139 Bengal infections in rabbit intestinal mucosa
    Amin A, Ali A, Kurunathan S, Cheong TG, Al-Jashamy KA, Jaafar H, et al.
    Histol Histopathol, 2009 05;24(5):559-65.
    PMID: 19283664 DOI: 10.14670/HH-24.559
    Vibrio cholerae is the causative agent of the infectious disease, cholera. The bacteria adhere to the mucosal membrane and release cholera toxin, leading to watery diarrhea. There are >100 serovars of V. cholerae, but the O1 and O139 serovars are the main causative agents of cholera. The present study aimed to compare the severity of intestinal mucosal infection caused by O1 El Tor and O139 V. cholerae in a rabbit ileal loop model. The results showed that although the fluid accumulation was similar in the loops inoculated with O1 and O139 V. cholerae, the presence of blood was detected only in the loops inoculated with the O139 serovar. Serosal hemorrhage was confirmed by histopathological examination and the loops inoculated with O139 showed massive destruction of villi and loss of intestinal glands. The submucosa and muscularis mucosa of the ileum showed the presence of edema with congested blood vessels, while severe hemorrhage was seen in the muscularis propria layer. The loops inoculated with O1 El Tor showed only minimal damage, with intact intestinal villi and glands. Diffuse colonies of the O139 serovar were seen to have infiltrated deep into the submucosal layer of the intestine. Although the infection caused by the O1 serovar was focal and invasive, it was more superficial than that due to O139, and involved only the villi. These observations were confirmed by immunostaining with O1 and O139 V. cholerae-specific monoclonal antibodies. The peroxidase reaction demonstrated involvement of tissues down to the submucosal layer in O139 V. cholerae infection, while in O1 El Tor infection, the reaction was confined mainly to the villi, and was greatly reduced in the submucosal region. This is the first reported study to clearly demonstrate the histopathological differences between infections caused by the O139 Bengal and O1 El Tor pathogenic serovars of V. cholerae.
    Matched MeSH terms: Immunoenzyme Techniques
  7. Factors associated with HER2 overexpression in breast cancer: Experience in an Asian developing country
    Tan GH, Choo WY, Taib NA, Yip CH
    Asian Pac J Cancer Prev, 2009;10(5):837-40.
    PMID: 20104975
    INTRODUCTION: The HER2 gene is amplified in up to 30% of human breast cancers, leading to overexpression of the HER2 protein on the cell surface. Overexpression of HER2 is associated with a more aggressive cancer and hence a poorer overall survival.

    OBJECTIVE: To evaluate the association between clinico-pathological features and HER2 overexpression in breast cancer.

    METHODS: This is a retrospective study conducted in the Department of Surgery, University Malaya Medical Centre. The association between HER2 overexpression, determined by immunohistochemistry, and other clinicopathological factors was evaluated in 996 patients with newly diagnosed breast cancer treated from 2005 to 2007 using univariate and multivariate logistic regression.

    RESULTS: HER2 overexpression occurred in 30.3% of patients. On bivariate analysis, HER2 overexpression was inversely related to ER expression (p<0.01) and PR expression (p<0.01). This overexpression was associated with a higher tumour grade, lymphovascular positivity and infiltrating ductal carcinoma subtype. On multivariate analysis, HER2 overexpression was significantly associated with higher tumour grade (p= 0.018, CI 1.25-11.04), PR negativity (p= 0.002, CI 0.30-0.77) and lymphovascular positivity (p= 0.042, CI 1.01-2.12).

    CONCLUSIONS: HER2 overexpression was observed in 30.3% of Malaysian female breast cancer patients. This group of patients represents a more aggressive subtype of breast cancer with higher tumour grade, PR negativity and lymphovascular positivity. No significant relationship was established between HER2 overexpression and age, race, lymph node, ER, pathology subtype and stage of disease from this study.

    Matched MeSH terms: Immunoenzyme Techniques
  8. Development of an indirect enzyme immunoassay using monoclonal antibodies for the measurement of 17alpha-hydroxyprogesterone in human serum
    Chong H, Cheah SH, Ragavan M, Johgalingam VT
    J Immunoassay Immunochem, 2009;30(2):166-79.
    PMID: 19330642 DOI: 10.1080/15321810902782863
    An indirect enzyme immunoassay for the measurement of total 17alpha-hydroxyprogesterone (17OHP) in serum using monoclonal antibodies generated in our laboratory was developed. Here, (a) instead of extraction with solvents, serum was heated to free protein-bound 17OHP and assay was performed at pH 9.6, (b) to ensure uniform assay conditions for both standards and samples, buffer for standards contained charcoal-stripped pre-heated pooled cord serum. Assays were done in 96-well EIA microplates pre-coated with 17alpha-hydroxyprogesterone-3-(o-carboxymethyl)oxime: bovine serum albumin. Secondary antibody was horseradish peroxidase-linked sheep anti-mouse IgG polyclonal antibody. The method was accurate and suitable for screening for congenital adrenal hyperplasia.
    Matched MeSH terms: Immunoenzyme Techniques*
  9. Fulltext Immunohistochemical expression of MAP1LC3A and MAP1LC3B protein in breast carcinoma tissues
    Othman EQ, Kaur G, Mutee AF, Muhammad TS, Tan ML
    J Clin Lab Anal, 2009;23(4):249-58.
    PMID: 19623642 DOI: 10.1002/jcla.20309
    Autophagy is a protein degradation process within the cell and its deregulation has been linked to various diseases and the formation of cancer. One of the important proteins involved in the autophagy process is microtubule-associated protein 1 light chain 3 (MAP1LC3). The aims of this study were to determine the MAP1LC3A and MAP1LC3B protein expression in both normal and cancer breast tissues and to determine the relationship between the expression of these proteins and type of tissues. Immunohistochemistry assessments were carried out on tissue microarrays consisting of breast tissues. MAP1LC3A expression was detected in 52/56 of normal breast tissue cores and 65/67 of breast cancer tissue cores. MAP1LC3B expression was detected in 55/56 of normal breast tissue cores and 67/67 of breast cancer tissue cores. MAP1LC3A and MAP1LC3B protein are expressed in the majority of normal and cancer breast tissues. A large number of MAP1LC3A and MAP1LC3B positive breast cancer tissues cores have high proportion of stained cells (81-100%) as compared with normal breast tissues. However, a significantly higher number of breast cancer tissues were found to express the MAP1LC3A protein with strong immunoreactivity as compared with the normal tissues, suggesting that MAP1LC3A may play a role in breast cancer development.
    Matched MeSH terms: Immunoenzyme Techniques/methods*
  10. Clinical relevance of CD10, BCL-6 and multiple myeloma-1 expression in diffuse large B-cell lymphomas in Malaysia
    Peh SC, Gan GG, Lee LK, Eow GI
    Pathol. Int., 2008 Sep;58(9):572-9.
    PMID: 18801072 DOI: 10.1111/j.1440-1827.2008.02273.x
    Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma, and it is recognized to constitute a heterogenous group of neoplasms. It can be divided into germinal center B-cell-like (GCB) and non-GCB subgroups. The aim of the present study was to evaluate the utility of immunophenotype subgrouping of DLBCL in a cohort of multi-ethnic Asian patients. A total of 84 reconfirmed de novo DLBCL were immunostained for the expression of CD10, BCL-2, BCL-6 and multiple myeloma-1. Thirty-three (39.3%) had the GCB phenotype, and the remainder (60.7%), the non-GCB phenotype. The results concur with most reports using a similar method of stratification. Forty-five patients had complete demographic and phenotype studies and 42 patients did not have rituximab treatment and had sufficient data for survival rate analysis. Similar to other studies, patients with combined low and low-intermediate International Prognostic Index score had better overall survival (P = 0.006). But patients with GCB phenotype did not have better prognosis, and BCL-2 expression was not associated with better prognosis. The expression of BCL-6 was associated with lower overall survival rate (P = 0.038). No apparent difference in overall and disease-free survival was noted between patients with GCB and non-GCB disease. BCL-6 expression by tumor cells appears to be associated with poorer prognosis.
    Matched MeSH terms: Immunoenzyme Techniques
  11. Fulltext CD44 expression and axillary lymph node metastasis in infiltrating ductal carcinoma of the breast
    Looi LM, Cheah PL, Zhao W, Ng MH, Yip CH
    Malays J Pathol, 2006 Dec;28(2):83-6.
    PMID: 18376796 MyJurnal
    Metastasising ability connotes one of the most important life-threatening properties of malignant neoplasms. Recent studies indicate that CD44 proteins, multifunctional cell adhesion molecules which contribute to "homing" of lymphocytes to lymph nodes as well as cell-cell and cell-matrix interactions, are potential markers of tumour progression. However, whether CD44 expression by human tumours contribute to increased metastatic risk remains controversial. In an attempt to clarify its role in breast cancer, we have investigated the correlation between CD44 expression by breast carcinoma and the presence of axillary lymph node metastases. CD44 expression was detected using a standard immunoperoxidase method on formalin-fixed, paraffin-embedded, primary infiltrating ductal breast carcinoma tissues taken from 60 female patients who underwent mastectomy with axillary node clearance. Tumours were graded according to the modified Bloom and Richardson criteria. 62% of patients had histologically-proven lymph node metastasis. 40% of primary cancers exhibited cytoplasmic membrane immunopositivity for CD44. 46% of primary tumours which have metastasied to axillary lymph nodes were CD44 positive whereas 30% of tumours which have not metastasised expressed CD44. CD44 positivity was expressed by 20% of grade 1, 31% grade 2 and 58% grade 3 tumours. Our results suggest that CD44 may have a role in the progression of breast cancer and emphasise the need to investigate its interaction with other mechanisms of cancer advancement.
    Matched MeSH terms: Immunoenzyme Techniques
  12. Fulltext Proliferating cell nuclear antigen (PCNA) activity in hepatocellular carcinoma, benign peri-neoplastic and normal liver
    Mun KS, Cheah PL, Baharudin NB, Looi LM
    Malays J Pathol, 2006 Dec;28(2):73-7.
    PMID: 18376794 MyJurnal
    Hepatocellular carcinoma (HCC) is among the ten most common cancers in Malaysian males. As cellular proliferation is an important feature of malignant transformation, we studied the proliferation pattern of normal and benign perineoplastic liver versus hepatocellular carcinoma in an attempt to further understand the tumour transformation process. 39 HCC (21 with accompanying and 18 without cirrhosis) histologically diagnosed at the Department of Pathology, University of Malaya Medical Centre between January 1992 and December 2003 were immunohistochemically studied using a monoclonal antibody to PCNA (Clone PC10: Dako). 20 livers from cases who had succumbed to traumatic injuries served as normal liver controls (NL). PCNA labeling index (PCNA-LI) was determined by counting the number of immunopositive cells in 1000 contiguous HCC, benign cirrhotic perineoplastic liver (BLC), benign perineoplastic non-cirrhotic (BLNC) and NL cells and conversion to a percentage. The PCNA-LI was also expressed as Ojanguren et al's grades. PCNA was expressed in 10% NL, 38.9% BLNC, 76.2% BLC and 71.8% HCC with BLNC, BLC and HCC showing significantly increased (p < 0.05) number of cases which expressed PCNA compared with NL. The number of BLC which expressed PCNA was also significantly increased compared with BLNC. PCNA-LI ranged from 0-2.0% (mean = 0.2%) in NL, 0-2.0% (mean = 0.3%) in BLNC, 0-3.6% (mean = 0.7%) in BLC and 0-53.8% (mean = 7.6%) in HCC with PCNA-LI significantly increased (p < 0.05) only in HCC compared with BLC, BLNC and NL. Accordingly, all NL, BLC and BLNC showed minimal (<5% cells being immunopositive) immunoreactivity on Ojanguren et al's grading system and only HCC demonstrated immunoreactivity which ranged up to grade 3 (75% of cells). From this study, there appears to be a generally increasing trend of proliferative activity from NL to BLNC to BLC and HCC. Nonetheless, BLNC and BLC, like NL, retained low PCNA-LI and only HCC had a significantly increased PCNA-LI compared with the benign categories. This is probably related to the malignant nature of HCC and may reflect the uncontrolled proliferation of the neoplastic hepatocytes.
    Matched MeSH terms: Immunoenzyme Techniques
  13. Fulltext Expression of survivin in fetal and adult normal tissues of rat
    Iskandar ZA, Al-Joudi FS
    Malays J Pathol, 2006 Dec;28(2):101-5.
    PMID: 18376799 MyJurnal
    Survivin is an inhibitor of apoptosis protein and regulates the cell cycle in the G2/M phase. Survivin is expressed during embryonic and fetal development, selectively over-expressed in common human cancers and completely down-regulated in normal adult tissue. This work was aimed at studying the expression of the survivin homologues and their subcellular distribution in fetal and normal adult tissues of rat. Survivin expression was evaluated by immunohistochemistry in formalin-fixed, paraffin-embedded tissue sections of fetal and normal adult tissues of rat using the polyclonal serum SUR12A-CFI. This serum demonstrated intense positive survivin staining in adult kidney, ovary and oviduct, and a variable expression in different fetal organs, with particularly intense expression detected in the adrenal gland, liver, stomach, small intestine, colon, kidney and skin. In both fetal and adult tissues, the expression was predominantly cytoplasmic. It was concluded that survivin was abundantly and prominently expressed during fetal development in rat and that the polyclonal anti-human survivin antibody SUR12A-CFI is reactive with rat survivin.
    Matched MeSH terms: Immunoenzyme Techniques
  14. Loss of human leucocyte antigen class I and gain of class II expression are early events in carcinogenesis: clues from a study of Barrett's oesophagus
    Rajendra S, Ackroyd R, Karim N, Mohan C, Ho JJ, Kutty MK
    J Clin Pathol, 2006 Sep;59(9):952-7.
    PMID: 16467164
    Human leucocyte antigen (HLA) expression is altered in oesophageal carcinomas compared with normal tissue. It is unclear, however, whether this phenotype precedes malignant transformation or results as a consequence of it.
    Matched MeSH terms: Immunoenzyme Techniques
  15. The prevalence of human cytomegalovirus seropositivity among blood donors at the Unit of Blood Transfusion Medicine, Hospital Universiti Sains Malaysia
    Ahmed SA, Al-Joudi FS, Zaidah AW, Roshan TM, Rapiaah M, Abdullah YM, et al.
    Southeast Asian J. Trop. Med. Public Health, 2006 Mar;37(2):294-6.
    PMID: 17124989
    Human cytomegalovirus (HCMV) is a species-specific DNA virus of the Herpetoviridae family. After a primary infection, HCMV persists in a latent form most probably in bone marrow progenitor cells or in peripheral blood monocytes. The virus can reactivate to result in shedding of the virus leading to virus dissemination and new infections. Immunocompromized patients are the ones most vulnerable to serious diseases occasionally acquired in blood transfusions. In a human population, HCMV seropositivity increases steadily with age to become approximately 100% in adults. This study was performed to detect seropositivity among regular blood donors in The Hospital of the Universiti Sains Malaysia, in the state of Kelantan. Using an enzyme immunoassay, it was found that 97.6% of blood donors were HCMV-positive. HCMV is highly prevalent and may be endemic in Kelantan. Hence, long-term strategies are required for the reduction of disease dissemination, and to prevent the exposure of immunocompromized patients to the virus.
    Matched MeSH terms: Immunoenzyme Techniques/methods
  16. Fulltext Plasma Total Homocysteine Concentration is Related To Insulin Level In Hypertensive Subjects
    Azizi Ayub, Awang Mat Zainal, Rafidah Hanim Mokhtar
    International Medical Journal Malaysia, 2006;5(1):1-13.
    MyJurnal
    Plasma total homocysteine levels (tHcy) is lowered by high insulin levels, and it can be elevated in insulin-resistant states. However, it is uncertain whether plasma tHcy and insulin or any components of the metabolic syndrome has any relationship among hypertensive individuals. In this study the tHcy and insulin concentrations were measured by enzyme immunoassay techniques in samples from 41 (27 male and 14 female) participants. Components of the metabolic syndrome (insulin resistance) profiles were also evaluated. The participants’ age ranged from 31 to 67 years (mean+SEM, 52.1±1.3 years), body mass index from 20.2 to 38.3 kg/m2 (27.2±0.7 kg/m2), plasma tHcy concentration from 6.9 to 16.2 μmol/L (11.0±0.4 μmol/L), and plasma insulin 3.0 to 16.6 μIU/mL (7.3±0.5 μIU/mL). A significant negative correlation was found between tHcy concentrations and insulin levels (r=-0.358, P=0.011), but not with other variables (P>0.05). In conclusion, the tHcy concentration is significantly related to plasma insulin in hypertensive subjects. tHcy concentrations were independent of the components of the metabolic syndrome and other risk factors of coronary heart disease in hypertensive subjects.
    Matched MeSH terms: Immunoenzyme Techniques
  17. Salmonellosis in persons infected with human immunodeficiency virus: a report of seven cases from Malaysia
    Puthucheary SD, Ng KP, Hafeez A, Raja NS, Hassan HH
    Southeast Asian J. Trop. Med. Public Health, 2004 Jun;35(2):361-5.
    PMID: 15691137
    Persons infected with human immunodeficiency virus (HIV) have an increased risk of salmonellosis when compared to the general population. We describe seven such patients with Salmonella bacteremia, of whom two had recurrent salmonellosis. In the latter two cases the infection was unusually severe, characterized by widespread infection, bacteremia and relapse, despite standard antimicrobial therapy. HIV-infected individuals will benefit from education on the source of Salmonella, mode of acquisition and prevention through safe food handling and food preparation practices. Because of the difficulty of eradicating Salmonella infection in patients with acquired immunodeficiency syndrome, long-term suppressive treatment with antimicrobials is warranted.
    Matched MeSH terms: Immunoenzyme Techniques
  18. Immunohistochemical localisation of RET and p53 mutant protein of thyroid lesions in a North-Eastern Malaysian population and its prognostic implications
    Omar E, Madhavan M, Othman NH
    Pathology, 2004 Apr;36(2):152-9.
    PMID: 15203751
    To investigate RET and p53 expression in local thyroid lesions, in order to shed light on the pathogenesis of papillary carcinoma and explain the high prevalence of this condition among the nodular hyperplasia (multi-nodular goitre) cases.
    Matched MeSH terms: Immunoenzyme Techniques
  19. Immunohistochemical expression of pi class glutathione S-transferase and alpha-fetoprotein in hepatocellular carcinoma and chronic liver disease
    Yusof YA, Yan KL, Hussain SN
    Anal. Quant. Cytol. Histol., 2003 Dec;25(6):332-8.
    PMID: 14714299
    To determine whether tumor marker pi glutathione transferase (GST-pi) is expressed in hepatocellular carcinoma (HCC) and other chronic liver diseases and to compare its expression with that of alpha-fetoprotein (AFP).
    Matched MeSH terms: Immunoenzyme Techniques
  20. Dual variant of Epstein-Barr virus in Hodgkin/Reed-Sternberg cells: single-cell PCR study on latent membrane protein-1 gene
    Kim LH, Peh SC, Poppema S
    Int J Cancer, 2003 Nov 1;107(2):250-5.
    PMID: 12949802
    Isolation of single cells permits analysis of DNA or RNA from individual cells among heterogeneous populations. This technique is particularly useful in the study of classical Hodgkin's lymphoma (cHL) due to the scarcity of H/RS tumor cells among large numbers of reactive leukocytes. In a previous study, we found a high frequency of dual LMP-1 variant (concurrent presence of deleted and nondeleted variants) in cHL from whole-tissue sections. For the present study, we applied a single-cell isolation technique to determine the LMP-1 oncogene variant in EBV-associated H/RS cells. Five cases of EBV-infected cHL, containing nondeleted (n=1), deleted (n=1) and dual infection (n=3) based on whole-tissue section analysis, were selected for study. Paraffin-embedded tissue sections were stained with antibody to LMP-1 and positively stained H/RS cells isolated using a semiautomated micromanipulator. Each isolated single cell was subjected to PCR for amplification of the LMP-1 gene flanking the 30 bp deletion region and Xho1 restriction site. Cases with either nondeleted variant or the deleted variant showed similar LMP-1 variant expression in isolated single H/RS cells. However, 1 of the 3 cases with dual variants showed only the deleted variant in H/RS cells. The other 2 cases showed mixed patterns of deleted, nondeleted and dual LMP-1 variants in isolated single H/RS cells. All cases showed loss of the Xho1 restriction site, with the exception of the case with nondeleted LMP-1. Results of single-H/RS cell analysis of the Xho1 restriction site concur with those of whole-tissue section amplification. A mixed pattern of LMP-1 variants was observed in isolated H/RS cells, and it is speculated that this is due to the accumulation of mutation and deletion events.
    Matched MeSH terms: Immunoenzyme Techniques
Related Terms
Filters
Contact Us

Please provide feedback to Administrator (afdal@afpm.org.my)

External Links