A multi-template multiplex PCR assay for hepatitis B virus and human ß-globin.

The Hepatitis B surface antigen (HBsAg) is the hallmark of HBV infection. Detection of antibodies to HBs and the core (ie, HBsAg and HBcAb) are primary serological algorithms in the laboratory diagnosis of HBV. Detection of HBsAg DNA is an important supplement to serological diagnosis especially in clinical cases. Simultaneous amplification of internal cellular controls is a good indicator of sample quality. Human ß-globin is a well characterized housekeeping gene (HKG) that is often applied as internal controls (IC) in molecular diagnosis. In this study, individual plasmid clones of the human ß-globin and HBs genes were constructed. These plasmid constructs have been applied to characterize a multiplex PCR assays for HBs and ß-globin genes. The findings suggest detection limits of less than 10 genome copies of either template In vitro using conventional and multiplex PCR conditions. Under the multiplex conditions, co-amplification of ß-globin and HBsAg DNA had a resultant effect on assay sensitivity. This study further highlights the importance of molecular diagnosis in HBV infectious individuals. If fully optimized, this assay could provide a possible diagnostic complement to serological detection in developing countries.

Human brucellosis in Northern Saudi Arabia.

OBJECTIVES: Analysis of the clinical features, laboratory findings, treatment given and complications seen in brucellosis patients at the Northern Area Armed Forces Hospital, Hafr Al-Batin, Kingdom of Saudi Arabia. METHODS: We retrieved and reviewed the record charts of all patients from January 1995 to December 2001 with a clinical diagnosis of brucellosis whose brucella agglutination titre was 1:160 or greater from the Medical Records Department of Northern Area Armed Forces Hospital, Hafr Al-Batin. We extracted from the files the information on age, gender, occupation, history of raw milk or milk products ingestion, presenting symptoms and physical signs. We also noted the results of routine laboratory tests, treatment given, outcome of treatment and complications. RESULTS: One hundred and fifty-nine patients (males 101, females 58 with a ratio of 1.7:1) had a diagnostic label of brucellosis and a brucella titre of > or -1:160. Thirty-three (20.8%) were < or -12, 96 (60.3%) were 13-40 years old. Twenty-six (16.4%) were 14-60 years while 4 patients (2.5%) were > or -60 years. Fever (> or -=37.7 degrees C) featured in 126 (79.2%) patients; joint pain in 112 (70.4%); while 77 (48.4%) had bone pain. We recorded the abdominal pain in 18 patients (11.3%) vomiting in 9 (5.7%) and anorexia in 6 (3.8%); splenomegaly in 6 (3.8)%, hepatomegaly and lymphadenopathy in 2 (1.3%) patients. Brucella tube agglutination titres ranged from 1:160 to 1:5120. Thirty-eight (35.8%) patients had anemia (Hb <12 gms/dl); 12 patients (9.8%) had lymphocytosis (lymphocyte count >1 k/L). Ten patients (6.2%) had bacteremia. We used Rifampicin and doxycycline in 87 cases (54.7%), doxycycline and streptomycin in 33 (20.8%), and rifampicin and streptomycin in 20 (12.6%) for 6 weeks or longer (we used combinations including septrin in the remaining patients). We recorded relapse in 18 patients (11.3%). Pneumonia, epididymo-orchitis in 2 cases (1.3%) each, abortion, threatened abortion in one case each, complicated the disease in these patients. CONCLUSION: Brucellosis is endemic in Northern Saudi Arabia as in other parts of the Kingdom. The clinical and laboratory features and response to therapy are also similar.

Sickle cell anemia morbidity in Northern Saudi Arabia.

OBJECTIVE: Documentation of the morbidity in sickle cell patients presenting at the Northern Area Armed Forces Hospital, Hafr Al Batin, Kingdom of Saudi Arabia and the burden of delivering care to these patients. METHODS: The record charts of all Saudi patients with Sickle Cell Anemia (SCA) admitted between June 1994 through to July 2001 were retrospectively analyzed. Focus was on age of patients, frequency of admissions, duration of hospital stay and requirements for blood transfusions. RESULTS: The study embraced a total of 84 patients (53 males and 31 females) giving a male to female ratio of 1.7:1. The mean age of patients was 10.8 years while the mean age at first admission was 3.8 years. The frequency of admissions peaked at 754 admissions (51%) in the age group 8-15 years and 326 (22%) admissions in the age group 4-8 years. The most frequent causes for admissions were in order of frequency, pain crisis, hemolysis, infections and anemia. Maximum hospitalization occurred in the age group 12-18 years. In 415 occasions admission of patient lasted for only one day; in 470 instances for 2 days and in 166 patients for 3 days. These gave a total of 3945 patient days of which 1255 days (31.8%) were for 2 days or less. CONCLUSION: Many of our patients were admitted for very short periods of stay in the ward for pain control, rehydration or blood transfusions. Costs can be reduced by managing them in a short stay ward. It would be helpful if a National Guideline for Management of Patients with SCA can be drawn up by a Committee of Clinical Hematologists and Clinicians with wide experience in management of patients with hemoglobinopathies. Such guidelines will aid Internists and Primary Health Care Physicians in smaller hospitals in the management of patients with SCA.