Method Verification of the Caretium XC-A30 Automated Erythrocyte Sedimentation Rate Analyser for Erythrocyte Sedimentation Rate.

Background: The erythrocyte sedimentation rate (ESR) analyser is widely used in haematological testing. In addition to the Westergren method, new automatic methods for ESR measurements have been developed. We aimed to study the reliability, precision, accuracy and stability of the Caretium XC-A30 automated ESR analyser. Methods: Ethylenediamine tetraacetic acid (EDTA)-treated blood samples were analysed via the Caretium XC-A30 automated ESR analyser and the Westergren method to compare accuracy. Precision was assessed using control samples and patient samples were classified into three groups-low, medium and high-according to their rates of sedimentation. Moreover, a stability test was performed. Results: The correlation coefficient of the results of the Caretium XC-A30 and Westergren analyses was 0.97. The correlation coefficient of ESR values obtained from the two methods assessed in the low, medium and high groups were r = 0.80, r = 0.68 and r = 0.74, respectively. The coefficient of variation of within-run (%CVw) and between-run (%CVb), with replicates performed with commercial controls samples, were 7.54% and 8.04% for the normal control and 4.68% and 3.50% for abnormal control, respectively. The %CVw obtained with patient samples in the low, medium and high groups were 10.68%, 13.13% and 4.45%, respectively. The Caretium XC-A30 measurements were stable for up to 24 h when samples were stored at 4 °C. Conclusion: The Caretium XC-A30 ESR analyser proved to be a suitable instrument for routine analysis of ESR.

Prenatal diagnosis of α- and ß-thalassemias in southern Thailand.

We report our work on prenatal diagnosis of α-thalassemia, ß-thalassemia and other hemoglobinopathies in southern Thailand. DNA-based diagnosis was offered to 1906 pregnancies at risk for thalassemia using a combination method of multiplex-PCR and reverse dot blot analysis to detect seven α-globin and 47 ß-globin mutations. The most commonly detected mutation of α0-thalassemia was a South-East Asian deletion (98%), followed by a Thai deletion (2%). Twenty-eight ß-globin mutations were identified. Fourteen common mutations, including cod 19 A-G (18.6%), cod 41/42 -TCTT (14.4%), IVS1#5 G-C (13.2%), 3.5 kb deletion (9.2%), cod 17 A-T (7.7%), -28 A-G (7.3%), IVSI#1 G-T (7.1%), 12.5 kb deletion (δß)0 (5.7%), HPFH6 (4.2%), IVS2#654 C-T (2.7%), 45 kb deletion (1.9%), Asian Indian-inversion-deletion Gγ(Aγδß)0 (1.6%), cod 41 -C (1.5%) and cod 71/72 +A (1.3%) were detected, accounting for 96.5% of all mutations detected. The most common hemoglobin variant was Hb E, accounting for 97.86%. Prenatal diagnosis of 1906 couples at risk showed 22.0% normal, 51.2% carrier and 26.8% affected. The present study provides important information for diagnosis and control of severe thalassemia diseases.

Molecular basis and hematological features of hemoglobin variants in Southern Thailand.

Hemoglobinopathy (abnormal hemoglobin or hemoglobin variant) is an inherited disorder that results in the abnormal structure of globin chains of the hemoglobin (Hb) molecule. Many abnormal Hbs have been characterized worldwide, including more than 20 variants in Thailand. The Bio-Rad Variant II HPLC system is used for investigating hemoglobin variants at Songklanagarind Hospital. This system has been shown to be a sensitive, specific, and reproducible method, but some hemoglobin variants such as Hb Tak and Hb D-Punjab cannot, as yet, be clearly separated by this method. The aim of this study was to investigate the prevalence of hemoglobinopathy in southern Thailand using DNA sequencing and study the severity of each hemoglobin variant. A total of 58 hemoglobin variant samples were obtained from blood samples undergoing routine hemoglobin typing at Songklanagarind Hospital. Genomic DNAs were extracted from the samples, and the globin genes were analyzed by using PCR-direct sequencing. The molecular analysis revealed eight hemoglobin variants: 28 Hb C, 12 Hb D-Punjab, 7 Hb Tak, 4 Hb G-Makassar, 2 Hb Lepore-Hollandia, 2 Hb Q-Thailand, 2 Hb O-Indonesia, and 1 Hb Hope. The distribution of hemoglobin variants in southern Thailand is associated with geographic and/or ethnic backgrounds. This study may help hematologists understand better the prevalence of hemoglobin variants and their hematological features in this region.