Application of DNA Microarray in Genetic Mutation Detection in Patients with Thalassemia / 中国实验血液学杂志

OBJECTIVE@#To perform dried blood spots thalassemia gene detection in patients with positive blood phenotypes by microarray technology, and evaluate its value in clinical detection.@*METHODS@#DNA samples were extracted from dried blood spots of 410 patients. Microarray technology was used to detect 3 deletion and 3 non-deletion types of α-thalassemia and 19 β-thalassemia point mutations which were common gene mutions in China.@*RESULTS@#There were 357 positive cases in all the 410 tested samples with the positive rate 87.07%, among which 299 cases (72.93%) carried deletion or point mutations of α-thalassemia, 29 cases (7.07%) carried point mutations of β-thalassemia and 29 cases (7.07%) carried gene mutations of complex αβ-thalassemia syndrome. The mutations of α-thalassemia were involved with --@*CONCLUSION@#The most common genetic mutations are --

Methodological Evaluation of Microarray in the Detection of α-Thalassemia / 中国实验血液学杂志

OBJECTIVE@#To proceed the clinical evaluation of DNA microarray for thalassemia gene detection.@*METHODS@#Peripheral blood samples of 166 thalassemia gene test subjects were collected and tested for thalassemia genes by microarray chip method and Gap-PCR method combined with PCR-reverse dot blot hybridization method according to double-blind control test. The specificity, sensitivity, positive predictive value, negative predictive value, and total coincidence rate of the microarray chip method were evaluated. When the two methods were inconsistent, multiplex ligation dependent probe amplification (MLPA) was used to verify the deletional α-thalassemia.@*RESULTS@#Compared with Gap-PCR method, specificity, sensitivity, positive predictive value, negative predictive value, Youden index, and total coincidence rate of microarray chip method was 100% (70/70), 96.88% (93/96), 100% (93/93), 95.89% (70/73), 0.969, and 97.59% (162/166), respectively, while compared with PCR-reverse dot blot hybridization method was 100% (125/125), 100% (41/41), 100% (41/41), 100% (125/125), 1, and 100% (166/166), respectively.@*CONCLUSION@#The microarray chip method for α-thalassemia gene detection shows the advantages of high specificity, sensitivity, and throughput.

Evaluation of detection of Epstein-Barr virus Rta/IgG in nasopharyngeal carcinoma / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the clinical value of Epstein-Barr virus (EBV) Rta/IgG in the diagnosis of nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Serum samples derived from 211 untreated patients with NPC, 413 subjects including 203 non-NPC ENT patients and 210 healthy volunteers as control were examined for the presence of antibodies directed against Rta/IgG by using enzyme-linked immnunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve was applied to perform methodical evaluation of this tumor marker.</p><p><b>RESULTS</b>The rA value median of Rta/IgG in NPC group was significantly higher than one in control group (P < 0.001). The area under ROC was 0.933. The sensitivity and specificity of this marker were 90.5% and 90.1%, respectively, when the best cutoff value was defined.</p><p><b>CONCLUSION</b>Rta/IgG detected with ELISA method is a new target of EBV, and may be one of important marker for NPC diagnosis.</p>

Evaluation of combined determinations of Epstein-Barr virus antibodies for nasopharyngeal carcinoma assessed with receiver operating characteristic curve based on logistic regression / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the diagnostic value of combined determination of Epstein-Barr virus (EBV) antibodies for nasopharyngeal carcinoma (NPC), including immunoglobulin (Ig) A against EBV capsid antigens (VCA), IgA against early antigens (EA), IgG against BRLF1 transcription activator (Rta) and IgA against EBV nuclear antigen-1 (EBNA1), assessed with receiver operating characteristic (ROC) curve based on logistic regression.</p><p><b>METHODS</b>Serum samples derived from 211 untreated patients with NPC and 203 non-NPC ENT patients were examined for the presence of VCA/IgA and EA/IgA by immunoenzymatic assay, Rta/IgG and EBNA1/IgA by enzyme-linked immnunosorbent assay (ELISA). The different Logistic regression models were established for various combined determinations of antibodies, respectively. Using the predicted probability as the analyzed variable, ROC curve was applied to evaluate the diagnostic accuracy of different combined determinations.</p><p><b>RESULTS</b>The sensitivity of VCA/IgA (98.1%) and the specificity of EA/IgA (98.5%) were the highest while detecting solely. The results which were analyzed by ROC curve based on Logistic regression showed that the sensitivity and specificity were improved. In two-marker combinations, VCA/IgA + Rta/IgG whose area under ROC curve (AUC) was 0.991 had the highest diagnostic accuracy, and its sensitivity, specificity and Youden index were 94.8%, 98.0% and 0.928 respectively. No significant difference of AUC were found comparing VCA/IgA + Rta/IgG with VCA/IgA + Rta/IgG + EBNA1/IgA and four-marker combination( P > 0.05), of which sensitivity, specificity and Youden index were 94.8%, 98.5%, 0.933 and 96.7%, 97.0%, 0.937, respectively.</p><p><b>CONCLUSION</b>The approach of ROC curve based on Logistic regression can improve synthetic efficiency for combined determination of multiple markers. The combined determination of VCA/IgA and Rta/IgG with a complementary effect is optimal for NPC serodiagnosis.</p>