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Effective screening for a-thalassemia 1, b-thalassemia and Hb E at the Diagnostic Clinical Microscopy Unit, Srinagarind Hospital

According to the policy of  the Ministry of  Public Health, severe thalassemia diseases targeted for prevention and control include homozygous a - thalassemia 1 (Hb Bart’s hydrops fetalis), homozygous b - thalassemia and b - thalassemia / Hb E disease. The objective of this study is to evaluate the effectiveness of thalassemia screening using a combined MCV (cut off; 80 fl) and KKU-DCIP-Clear at the Diagnostic Clinical Microscopy Unit, Srinagarind Hospital, Faculty of Medicine, Khon Kaen University. Studied was carried out on 100 subjects. After screening, definite diagnosis of each case was done by standard Hb and DNA analyses. Thalassemia identified included 1 b - thalassemia trait, 1 a - thalassemia 1 trait, 1 Hb E trait with a - thalassemia 1 trait, 5 Hb E trait with a - thalassemia 2, 2 Hb E trait with Hb Con Sp trait, 1 Hb E trait with a - thalassemia 2 / Hb Con Sp, 24 Hb E trait, 3 Homozygous Hb E, 1 homozygous Hb E with Hb Pakse’ trait, 12 a - thalassemia 2 trait, 1 a - thalassemia 2 / Hb Con Sp , 1 Hb Lepore trait and 47 non-thalassemia. No false negative was found for screening of the three important thalassemia carriers but 6 false positive cases were encountered. Therefore, the sensitivity and specificity  of screening for a - thalassemia 1, b - thalassemia and Hb E using MCV and KKU-DCIP-Clear were 100 % and 90.2 %, respectively. The specificity was increased to 91.8 % when another red blood cell parameter, mean cell hemoglobin (MCH) (cut off; 27 pg) was used additionally. This study confirmed that the MCV and MCH values obtained from quality controlled electronic blood cell counter could be used for screening of a and b - thalassemias instead of the OF test. Using MCV and MCH in combination with DCIP test should provide effective screening for severe thalassemia carriers in routine practice.      

Proficiency testing in laboratory diagnosis of common thalassemia and hemoglobinopathies in Thailand

To improve the efficiency of laboratory diagnosis of common thalassemia and hemoglobinopathies in Thailand,the proficiency testing program was set up at the Centre for Research and Development of Medical DiagnosticLaboratories (CMDL), Khon Kaen University. The KKU-Hb controls were sent to laboratory members togetherwith essential hematological parameters. Each time, two control samples designated as husband and wife were sent.Upon receiving, all laboratory members analyzed control samples in their routine practices and interpreted the resultusing hematological data received and result of Hb analyses in their laboratories. The result of laboratory investigationsand the interpretations as well as the risks of having fetuses with 3 severe thalassemia diseases including homozygousα- thalassemia 1, homozygous β-thalassemia and β-thalassemia / Hb E disease were applied into the form providedand sent back to CMDL. Three cycles were investigated with 21, 23 and 66 participant laboratories, respectively.All control samples were received within appropriate times and conditions. It was found that more than 90 % ofparticipant laboratories could report acceptable levels of Hb A2 and Hb F and give accurate interpretation. Memberswere analyzed and grouped into 4 different quality groups;Excellent, Good, Fair and Need improvement.The proportions of members in the Excellent, Good, Fair and Need improvement groups were respectively found tobe (81.0, 9.5, 4.75 \& 4.75 %) in the first cycle and (69.6, 0, 21.7 \& 8.7 %) in the second cycle and (56.0, 18.2,24.3 \& 1.5 %) in the third cycle. It was found that the values of Hb A2 and Hb F were reported quite accuratelyfrom each laboratory member. However, when samples with complicated data were supplied, the increased inmis-interpretation and evaluation of relative risks were observed. This result indicates the requirement of furtherimprovement in the laboratory interpretation and knowledge related to laboratory diagnosis of thalassemiaand hemoglobinopathies of the participants. The evaluation system developed should prove useful in both developmentof external quality control program in laboratory diagnosis and further facilitate the prevention and control programof thalassemia and hemoglobinopathies in Thailand.

Apolipoprotein E polymorphism and dyslipidemia in Thais

Apolipoprotein E (apoE) is a constituent on lipoprotein surface and plays an important role in lipid metabolism. There are three common isoforms of human apoE, designed apoE2, apoE3 and apoE4 that are coded by three polymorphic alleles of the APOE genes, e2, e3 and e4. Polymorphism of APOE influences the blood lipid concentration and may contribute to susceptibility to dyslipidemia. The present study was therefore to investigate the role of APOE polymorphism on blood lipid levels in Thai individuals. A total of 121 normolipidemic and 125 dyslipidemic subjects were recruited. DNA was isolated from peripheral blood leukocytes and APOE genotypes were determined by PCR-RFLP. Allele frequencies of e2, e3 and e4  were 9.3%, 77.8% and 12.8%, respectively. The respective prevalence of APOE genotypes for e2/e2, e3/e3, e4/e4, e2/e3, e2/e4 and e3/e4 were 1.2%, 62.2%, 0.8%, 11.8%, 4.5% and 19.5%. The frequencies of APOE allele and genotype of both groups as well as the relation of APOE genotypes to the risk of dyslipidemia were not significantly different. The elevation of some lipid parameters observed in male subjects and the higher level of lipid profile in dyslipidemic group may be due to other factors and gene polymorphisms.

Assessment of quality control producy for thalassemia screening using MCV and MCH approaches

A combined MCV/DCIP or MCH/DCIP is one of screening approach for thalassemia alternative to the OF/DCIP. Based on this approach, the MCV and MCH values obtained from electronic blood cell counters are of important that should be accurate and comparable. To evaluate the accuracy of thalassemia screening using the MCV and MCH measured from different instruments and settings, two quality control (QC) products, the RBC_A (MCV \> 80 fl and MCH \> 27 pg) and RBC_B (MCV \< 80 fl and MCH \< 27 pg), were prepared and sent to 13 participated laboratories. Rbc parameters of each QC product were measured routinely in each laboratory. By comparing to the initial values obtained in our laboratory, it was found that all Rbc parameters of the two QC products were accurately measured with the coefficient of variation (% CV) of 2.09 - 9.76 % for RBC_A and 1.11 - 11.04 % for RBC_B. Regarding to the instruments, the % CV of most Rbc parameters were less than 2 % for both RBC_A and RBC_B. To evaluate the accuracy of the MCV and MCH in application to thalassemia screening, the MCV cutoff value of 80 fl and MCH cutoff value of 27 pg were applied. The RBC_A was treated as negative sample and the RBC_B was treated as positive sample. By using the MCV approach, false negative results were obtained form 2 laboratories. In contrast, neither false negative nor false positive result was found with the MCH approach, suggesting that MCH might be more appropriate for thalassemia screening than MCV. The results indicate that these QC products might be used as an internal QC to evaluate the accuracy of Rbc parameters and as an external QC to evaluate the performance of thalassemia screening using the MCV and MCH approaches of each laboratory setting.