Highly Sensitive Multiplex RT-PCR System for the Detection of all Common BCR-ABL Transcripts Associated with Different Leukemias

After the recent characterization of leukemia-specific DNA rearrangements, molecular methods have become primary tools for the diagnosis and monitoring of many hematological malignancies, due to their superior sensitivity and accuracy over other conventional methods. The BCR-ABL fusion transcripts resulting from the t[9; 22] translocation are distinct hallmarks of Philadelphia chromosome positive [Ph[+]] leukemias. There are clear associations between different isoforms of the BCR-ABL fusion protein and specific phenotypes of these leukemias. Each isoform also has a significant prognostic value and can be a critical indicator of the clinical outcome. In this study we have adopted, with modifications, a highly sensitive and specific method to screen simultaneously for the most frequent BCR-ABL fusion transcripts, namely, p210 [b3a2/b2a2], p190 [ela2] and p230 [e19a2]. A multiplex reverse-transcriptase polymerase chain reaction [RT-PCR] protocol with nested primer strategy for each of the above fusion transcripts was carefully optimized. RNA integrity, cDNA synthesis and PCR amplification were checked using internal control primers for the normal untranslocated BCR gene. Over 100 clinical samples were collected from hospitals in Amman between 2003 and 2005. This system was applied successfully on 100 clinical samples previously diagnosed as chronic myelogenous leukemia [CML] and acute lymphocytic leukemia [ALL]. RNA extracts from established leukemic cell lines carrying the translocations of interest were used as external positive controls. Representative PCR products were sequenced to verify the specificity of the amplification system. Our multiplexed nested RT-PCR assay provides a sensitive, accurate, time-saving and cost-effective diagnostic tool for the diagnosis and monitoring of patients with Ph [+] leukemias

Epidemiology and management of hypertension among Bedouins in northern Jordan

To estimate the magnitude of hypertension, and its levels of awareness and control of hypertension among a community of Jordanian aborigines in Northern Jordan; and to compare the study findings with findings from other Jordanian communities. A systematic sample of 50% of all households of Qafqafa, Northern Jordan was selected and all members of the selected households were included in the study at Qafqafa Health Center, Qafqafa, Jordan. A structured questionnaire was administered to all study participants and measurement of blood pressure was performed according to a preset criteria. Data on 366 subjects included in the sample were collected during the months of April and May 2002. A total of 40 subjects or 10.9% were suffering from hypertension. Prevalence rate of hypertension was significantly lower than the 16.1% and 16.3% prevalence rates reported from other more modernized Jordanian communities. Logistic regression analysis indicated that hypertension was positively associated with masculinity, age, illiteracy, obesity, and with positive family history of hypertension. No association was detected between hypertension and each of smoking, Diabetes mellitus, and total serum cholesterol. This study showed that one-half [47.5%] of hypertensives were unaware of their diagnosis and more than one-half [57.1%] of those aware of their diagnosis did not achieve control of their hypertension. Population of the study community has significantly lower levels of hypertension than both semi-urban and recently urbanized Jordanian communities, and much of the rise in blood pressure levels is attributed to environmental factors. Moreover, hypertension management programs in Jordan are far below the optimal standards