HIV Prevention and Test Methodology Study Population: The study population included males and females with febrile illness that visited the above mentioned hospital.
Sample Size: Eighteen (18) blood samples was collected for the study.
Material: New unopened test packet, Alcohol swab, Sterile disposable lancets, Assay diluents. Timer, Sharps box, Pencil or pen, plastic syringe (5ml or 10ml), Disposal glove, Test tubes, EDTA bottles, Medical cotton wool, Spirit (70% ethanol), Standard micropipette, Microscope slide, Cover slide, microscope
Samples Collection: A sterile dry, preferably plastic syringe of the capacity required was selected e.g 2.5ml 5ml or 10ml. a soft tubing tourniquet or Valero fastening arm band was applied to the upper arm of the patient to enable the vein seen and felt. Using the index figure, felt a suitable vein selecting a sufficient large straight vein that does not roll with a direction that can be felt.
The punctured site was cleansed with 70% ethanol and allowed to dry. Using the thumb of the left hand holding down the skin below the puncture site, I made a vene puncture with the bevel of the needle directed upwards in the line of the vein. Steadily withdrew the plunger of the syringe at the speed it is taking the vein to fill.
I avoided movement of the needle to the vein. When sufficient blood was collected I released the tourniquet and instructed the patient to open his or her fist. Removed the needle and immediately pressed on the puncture site with a piece of dry cotton wool. I removed the tourniquet completely. The needle was carefully form the syringe and filled the container with required volume of blood.
The needle was discarded. The blood was immediately mixed with ethylene diamine tetra acetate (EDTA) and lablled accordingly.
HIV screening: Method of Serum Preparation (HIV Prevention)
: Exactly 2ml of blood sample was collected from patient and introduced into sterile dry tube and was allowed to clot and refract which it 1 centrifuged at 3500 rpm for 5 minutes to give clear sera which was stored at 20oc and analyzed within 2-3 days of collection (Nwuzo et al., 2009).
Examination of blood for HIV: The HIV status of the patients was determined by enzyme linked immunosorbent assay (ELISA) method using commercial available ELIS kits: ACON Diagnostics, USA, Biosystem, USA) and was confirmed by a second stage confirmatory test of two to three rapid tests with different principles (Capillus HIV ½ Assay, Trinity biotech, Ireland and Determines kits, Japan Co. LTD) of antibodies and antigen methods as recommended by WHO for low resource countries including Nigeria (CDC, 2004).
Microscopy Thin film preparation: A drop of flesh blood was gently made to touch one end of a clean grease free slide. A spreader was used to spread the blood along the glass slide. The spreader was held at suitable angle 45 and pushed the spreader along side, drew smeared forming a thin film. The thin film was allowed to air dry, then fixed with three drops of methanol. The forceps was used to place the slide on a staining trough. Three percent (3%) Giemsa solution was prepared buffer or distilled water in sufficient quantity to fill the number of staining trough. The stain was allowed to stain for 30-45 minutes out of sunlight. Clean water was poured gently on the surface of glass slide was gently poured off. The forceps was used to remove the slide and placed it on the slide cleaned and examined with a microsocope.
Thick blood film preparation: A fresh drop of blood was gently touched onto the centre of a clean grease free slide. A beveled piece of glass (spreader) a little narrower than the slide was used to spread a drop of blood in a circular form.
The thick film was allowed to air dry thoroughly; it was not fixed so as to permit dehaemoglobinization. The forceps was used to place the slide on a staining trough. Three percent (3%) Giemsa solution was prepared in buffer or distilled water in sufficient quantity to fill the number of staining trough. The stain was poured gently into the staining trough, until all the slides were completely covered.
The stain was allowed to dry for 30-45minutes. Clean water was poured gently on the surface to remove staining solution deposit. The remaining water on the surface of the glass slide was gently poured off. The forceps was used to remove the slide and placed on the slide rack to drain off and allowed to air dry at room temperature. The back of the slide was cleaned and examined with a microscope.
Statistical Analysis: The density of the malaria in HIV negative and HIV positive individuals was calculated and their difference in proportion was evaluated using tables.
|TABLE I. CHARACTERISTICS OF CHILDREN WITH HIV MONO-INFECTION AND THOSE WITH CO-INFECTION|
|CharacteristicsGender (N(%))FemaleMale Mean age (yrs) (SD)||HIV only(N=135)55(40.7)80(59.3) 6.69 (4.82)||HIV/HCV(N=8)4(50.0)4(50.0) 7.48 (3.91)||Total6491||p-value0.81 0.83|
|Route of acquisition of HIV (N (%))VerticalInfected bloodSexual Unknown||111(82.2)9(6.7) 2(1.5) 13(9.6)||7(87.5)0(0.0) 0(0.0) 1(12.5)||12711 2 15||0.76|
|Socio-Economic Status (N(%))HighMiddleLow||25 (26.0)17 (17.7) 54 (56.3)||0 (0.0)0 (0.0) 3 (100.0)||2618 61||0.30|
|WHO clinical stage at entry* (N(%))Stage I (A1 & P1)Stage II (A2 & )||34(25.2)37(27.4)58(43.0) 6(4.4)||1(12.5)2(25.0)5(62.5) 0(0.0)||394167 8||0.63|
|TABLE II. ASSOCIATION OF HEPATTIS B AND C CO-INFECTED WITH RISK FACTORS FOR HORIZONTAL TRANSMISSION|
|Risk factorCircumcision (N(%)YesNo||Co-infected 13(65.0)7(35.0)||Mono-infected 98(73.1)36(26.9)||p-value 0.44|
|Ear piercing (N(%))YesNo||9(45.0)11(55.0)||61(45.2)74(54.8)||1.00|
|Unsafe injections (N(%))YesNo||2(10.5)17(89.5)||49(37.4)82(62.6)||0.74|
|Blood transfusion (N(%))YesNo||4(22.2)14(77.8)||49(37.4)82(62.6)||0.30|