Effect of Blood Bags Storage Time on The Quality of HIV Pre Transfusion Test Result: Preliminary Study in Blood Bank’s dr.

Research on the effect of storage time on blood quality has been conducted. The purpose of this study is to determine the quality changes caused by the length of storage. The test parameter is the change in blood to become HIV positive. The blood testing process is carried out by immune chromatography on blood that has been stored for 2 weeks, 3 weeks, 4 weeks and 5 weeks. Test results show changes in blood quality during storage. Time does not show a difference in changes in blood quality, however it needs to be aware, because the stored blood changes


INTRODUCTION
Blood transfusion is the process of taking blood from someone voluntarily to be stored in a blood bank and then used in blood transfusions by medical personnel [17].Blood transfusion is a therapeutic procedure in which it functions as a healing, but contaminated blood transfusion can transmit infectious diseases and can endanger life rather than save lives].Safe blood transfusion services are the foundation of an effective health care system [2].Blood for transfusion is considered safe when it is: 1) Donated by a carefully selected, healthy donor; 2) Free from infections that could be harmful to the recipient; 3) Processed by reliable methods of testing, component production, storage and transportation; and 4) Transfused only upon need and for the patient's health and wellbeing [18].
One of the deadly diseases that are at risk of being transmitted through blood donation is HIV (Human Infectious Disease).Transmission of HIV through blood transfusion was first known in late 1981 and early 1983 [18].Blood banks have also begun to ask donors about various high-risk behaviors, even before HIV antibody screening was carried out, which in fact has been able to reduce the number of HIV infections transmitted through transfusions [3].In Indonesia, East Nusa Tenggara is one of the areas with high HIV patients [19], so blood donor needs to be considered.An important aspect of managing blood for transfusion/donation is storage.According to the Naim report (2014) storage time affects the quality of platelets, erythrocytes and hemoglobin.
There is a real case of a kidney failure patient in East Nusa Tenggara who was HIV positive after receiving a blood transfusion.In 2015, a 38-year-old man was convicted of Kidney Failure at RSUD dr.Ben Mboi Manggarai District, East Nusa Tenggara Province, for 9 months recorded 47 blood bags that have been transfused to the man, after being transfused as many as 47 blood bags (whole blood) the man was convicted of HIV & AIDS.In 2017 there is a 23-year-old man who has been donated from him twice as long as the last 10 months, when he was about to donating the blood the third time he was convicted of having the HIV virus.Based on this fact it is necessary to test related to the effect of bag storage time on HIV test.

Tools and Materials
The tools used in this study include dropper pipettes, pin pliers, refrigerators, scissors, centrifuges, test tubes, test tube racks.While the research material consisted of Whole blood, sd rapid test, oncoprobe rapid test, inteck rapid test..

Research procedure
This research is an experimental study conducted at the Blood Bank of RSUD dr.Ben Mboi Ruteng.Manggarai Regency East Nusa Tenggara Province is preliminary to determine the effect of the time of storage of blood bags on blood quality, in this case seen from the presence or absence of the HIV virus in the blood.The design of this study is a Case Study (Case Study Design) that is by comparing the differences in the results of immuno chromatographic tests on blood bags (whole blood) that have been screened for HIV (free of HIV) by PMI, then stored in a refrigerator at a temperature of 2° C to 6 ° C. [6]; [20].The treatment given is storage time which is 2 weeks, 3 weeks, 4 weeks and 5 weeks.It is based on a modification of Naim's (2014) study and considers the incubation period of HIV [21].

Screening work procedures [4]:
➢ Prepare equipment and materials as well as PPE (Personal Protective Equipment) for re-screening ➢ Performed at room temperature, the test should be done in less than 20 minutes to get the best results.➢ Place the test kit on a flat, clean surface.The dropper pipette is held vertically and then drop 1 drop of serum or plasma (± 30-40 µl) into the specimen well (S), then add 2 drops of buffer.Avoid air bubbles.➢ Wait until the red line appears, the results should be read within 15 minutes.
Note: results are not interpreted after 20 minutes.

Interpretation of results
Positive (reactive): if a red line appears on the control line and the test line.Negative (non-reactive): Only one red line appears on the control, and invalid: no red line appears at all or appears only on the test (T) [4].

Data analysis
After the data is collected then the data is processed using data analysis with Kruskal Wallis test which is part of static non parametric which aims to assist researchers in distinguishing the performance results of groups contained in the sample into two groups with two different criteria [5].All immunochemical methods are based on a highly specific and sensitive reaction between an antigen and an antibody.Antigen is a substance that induces the production of antibodies [ie, proteins from the class of immunoglobulins (MW about 150 kDa) that are produced in the immune system of any vertebrate or human as a result of a defense reaction (immunity) to this for-eign substance [11].Antibodies are a large family of glycoproteins that share key structural and functional properties.Functionally, they can be characterized by their ability to bind both to antigens and specialized cells or proteins of the immune system.Assay specificity and sensitivity is determined by the quality of antibodies used in the method.Polyclonal antibodies are isolated from the serum of immunized animals, usually rabbits, and the serum is a combination of numerous antibodies with different specificities and affinities.Monoclonal antibodies are more homogenous in terms of specificity and affinity because of the production method involving immunization of mice followed by hybridoma technologies [11]; [12].

Immuno chromatography uses the principle of chemistry in its work
The results of the examination affect the storage period of blood bags (whole blood) on the results of pre-transfusion HIV tests using immunochromatography methods at Blood Bank RSUDdr.Ben Mboi Ruteng Manggarai Regency, NTT Province.The study was conducted with repeated measurements (repeated measure of anova) where the initial screening conducted by the PMI for 15 whole blood from the donor in a period of 1 week, 2 weeks, 3 weeks, 4 weeks, to 5 weeks, the following results were obtained:

DISCUSSION
Based on the table 1 that of the 15 blood bags (whole blood) there were found 2 who were reactively convicted of HIV.From the table 1, 1 is shows non-reactive and is shows reactive in each recipient 1-15, shows that at weeks 2 through 5 of the blood bag (whole blood) of the donor recepient 10 becomes reactive and the donor recepient becomes reactive from week 3 to week 5 among the people of Manggarai Regency, East Nusa Tenggara at the Blood Bank of RSUD dr.Ben Mboi Ruteng Manggarai Regency, NTT Province, which was initially non-reactive with HIV, has changed to become reactive to HIV after being re-screened for a period of 2 to 5 weeks in the blood transfusion unit.
The function of red cell storage is to maintain the functionality and viability of red cells throughout the approved storage period [15;16].The difficulty that is shared by modern storage mediums, however, is that red cell functionality and viability are progressively impaired during storage by three interrelated mechanisms: altered metabolism; increased oxidative stress; and membrane damageinitially interact with lipopolysaccharide, a highly anionic outer membrane, and glycolipid and then disrupt the membrane locally [5].
With the data quality results of the category (1-2) for the data analysis using nonparametric analysis, namely the kruskall walis method.Where, the critical test of the walis is a non-parametric test used to compare three or more groups of sample data.Based on kruskall walis test, known that no significant difference even the second and fifth weeks there are HIV reactive blood bags.
Changes can occur in the structure of stored blood [6,7], including the blood of donated HIV patients.This research shows that even though people who contain HIV are initially negative (incubation period), HIV will be active even if the blood is stored.In 4°C liquid storage, the biochemical and mechanical properties of red blood cells (RBCs) deteriorate progressively.When blood is stored in blood bank, biochemistry and physical properties of RBCs are altered because of storage conditions.These are referred to as storage lesions.Under normal conditions in the body's circulation, these do not occur as optimum temperature, pH, nutrient concentration and waste product removal are maintained [13;14].
Beside that storage has a negative effect on RBC oxygen delivery [8] and emerging evidence suggests that allogenic RBC infusion may actually harm some recipients.Considerable evidence suggests that transfusion increases the risk of serious complications and death in critically ill patients, especially in patients who are undergoing cardiac surgery.Current research indicated that the RBC hypothermic storage lesion is responsible for the association of blood transfusion with an increased length of stay in the hospital, impaired tissue oxygen use, proinflammatory and immunomodulatory effects, increased infections, multiple organ system failure, and ultimately increased morbidity and mortality [9].Clinical implications, collectively known as the RBC storage medium lesion, is in part related to bioreactive substances released by leucocytes in the storage medium, such as histamine, lipids, and cytokines, which may exert direct effect on metabolic and physical changes associated with the senescence, such as membrane reticulation, decrease in cell size, increase of cell density, alteration of cytoskeleton, enzymatic desilylation, and phosphatidylserine exposure, RBCs lose potassium 2,3-diphosphoglycerate (2, 3-DPG), Adenosine Triphosphate (ATP) stores, lipids and membrane, while becoming more rigid and demonstrating reduced oxygen offloading [10].

CONCLUSION
Blood storage time does not affect the results of immunochromatographic HIV testing.

Table 1 .
List of results of examination of the storage period of whole blood