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Human population has been affected numerous times by the deadly disease attacks, as was the case of the Black Plaque, affecting a major part of the population in the 14th century and a recent case of the SARS (severe acute respiratory syndrome) virus. The survival of these affected populations is primarily dependent upon the devising of effective cure for these new diseases.
But thankfully, because of the advancements in the technology, these types of diseases if had occurred in the recent times, would have been dealt with more effectively with minimum deaths. Technological advancements such as the molecular diagnostic techniques have equipped mankind with the skill of analysing and curing these infectious diseases much better. This has meant an increased assurance of mankind's survival as compared to the bleak survival chances hundred years ago.
hypothetical disease outbreak are defined by symptoms such as:
Flu like symptoms visible in patients.
These symptoms are coupled with anaemia, lethargy and severe bone pain.
After 4-6 weeks:
Recovering of patients from these symptoms without any symptoms present.
The other half of the population is signified by disease progression and a high fever remaining persistent leading to weight loss.
After 6-12 months:
Deaths of patients occur owing to the depressed immune system as a result of complications of infections.
Rarely visible in children before puberty and the middle aged or the elderly group.
The age group from 18-25 seen as the main victims.
Majority of the ethnic groups are affected except the Afro-Caribbean people.
Except the homosexual communities many bi-sexual and heterosexual communities are affected.
Possible disease mechanisms?
In the light of the occurring symptoms in the effected, one can further prod into the manners by which the disease manifests itself.
Firstly the most vital point is that this disease is caused by the bacterial and viral infectious agents. According to the available literature, majority of the patients recover from this disease before developing any severe and life threatening symptoms. Many patients are able to recover their health with the absence of any sort of long lasting effects. This is a positive sign indicating that the immune system of the body is able to respond and counteract certain portions of the virus or bacteria and thus was able to eliminate it completely.
The other remaining half of the patients, progress into the critical stage of the disease, by targeting the primary component of the body's immune system, the T-cell. The disease thus progresses into the highly metastatic lymphoma of the T-cell origin. Thus the defence mechanism of the body is compromised.
According to the epidemiological studies, the disease does demonstrate an unusual distribution pattern. In fact they are only occurring in certain groups depending on age and sex. The disease is sexually transmitted as studies show frequent occurrence of this disease among the heterosexual and bi-sexual people, while it is seldom found in the homosexual group.
So keeping in mind the above argument, the factors determining the occurring of the disease can be jotted down as:
The development of T-cell lymphoma is noticed in the affected patients after a time period of 12 months. The T-cell was discovered by Thomas Hodgkin in 1832, which was recognised as a significant component of the immune system. The T-cells were a particular group of the white blood cells that were known as the lymphocytes. The T-cells underwent reclassifications as new forms and sizes of the cells were discovered. The lymphoma re3ferred to in the modern study can therefore be a "Hodgkin's" or a "Non-Hodgkin's" lymphoma. In which the non-Hodgkin's lymphoma consisted of all the lymphoma that was not classified as a Hodgkin's ("The leukemia & lymphoma Society" May 2006).
One or more mutations do occur in one or more regulatory processes in order for the progression of the disease to such a high metastatic lymphoma in the patient affected. Disruption of cell cycle checkpoints and the disintegration of the DNA integrity genes are the processes occurring during such regulatory processes (Jacks et al., 2002.
The patients initially suffer from symptoms such as acute bone pain, lethargy and anaemia which then proceed further. To determine the appropriate cure would be possible by determining its location and the mutation mechanism taking place during these regulatory processes. Therefore it would be the best option to initiate the study with. The initial symptoms indicate that the disease is in a way developed via these symptoms or these symptoms may be the initiating factor behind the development of the life threatening T-cell lymphoma. The key factor in understanding the disease is to understand the preceding stages of the disease and hence to cure it at this stage. Therefore it would be necessary for patients to be treated as soon as possible with the development of the initial symptoms.
In the light of the above discussions, it is also noted that the patients suffering from the disease for 6 or more months, will have their T-cells being targeted. Therefore to devise an appropriate cure, it would be better to investigate and understand the manner in which these bacteria and viruses may attack the immune system of the patient.
DNA diagnostically testing
In order to gain knowledge of the disease it would be better to analyse the DNA forming this particular virus. The DNA reflects each and every characteristic, such as eye colour, personality, etc. Therefore the DNA of the virus will tell us about its attachment behaviour, development and reproduction if any. The study of the genetic make-up will enable unlocking the inner characteristics of the virus at the molecular level.
The mutations of two particular genetic types, the Oncogenes and the Tumor Suppressor Genes (TSGs), lead to cancers in people. Therefore analysis of the particular nucleotides of the patient's DNA by the usage of DNA diagnostic system will enable a comparative study which will prove beneficial for the study.
The example of human papilloma virus is a commonly occurring disease. It functions by assimilating its DNA with the host's DNA causing cervical cancer. The comparative study of the sample from an infected patient with that of a healthy individual will demonstrate the sequential differences occurring between the two.
There are other DNA systems available such as the Southern blotting system and the nucleic acid hybridisation. The nucleic acid hybridisation is the method in which the single strand of the DNA is attached to a particular membrane. A separate DNA probe is added which enables attachment. The target sequence is the first strand and the labelled strand is the second sequence which is added. The attachment of both the strands when occurs, will lead to detection with the help of suitable apparatus (Glick et al., 2003).
Cell culturing technique has been carried out since the mid 1900's and is still being practiced till today. Many laboratories are using this method to carry out diagnostic procedures by initiating with cell culture. The virus must however be placed in a culture to help in its growth so to enable the sufficient growth of infectious agents. The cell culture medium, for instance, used for the purpose of growth varies in composition according to the cell. The difference may be in terms of variable nutrients present. Hence the cell culture medium is thought carefully according to the cell.
After the completion of the first level, that is the selection of a growth medium, the cells are allowed to grow. After a certain level of growth is achieved the next step is initiated, which is the identification process. A range of tests are applied to determine the physiological properties making up the cell. The outcome is according to the make-up of the virus. The properties may include the required salt concentration, pH control, supply of gas, etc (Warrell et al., 2005).
The T-cell culture method is commercially used for research purposes owing to its usefulness. In spite of this there are many disadvantages to the process, such as:
Absence of growth.
Slow paced growth.
Equipment and maintenance cost (Zareen et al., 2009).
But certain viruses like the Chlamydia; the recognition of the properties is not possible. Thus this is a disadvantage of the process.
Polymerase Chain Reaction (PCR)
The polymerase chain reaction is frequently and widely used in various laboratories. The development of PCR was an achievement in itself because of its simple technique which can be utilised in the diagnostic laboratory. This technique can be conveniently modified to suit the researcher's requirements and is target specific. The technique is therefore a simplified and a straight forward approach towards analysing changes in the pathogen sequence.
The DNA fragments in a collection of template DNA is amplified, containing sequences not of interest. But to establish its effectiveness on a patient's sample, it must be seen that the RNA is utilised by many viruses to replicate and therefore lead to infections in the host. This is solved by the conversion of the RNA to a DNA sequence. The enzyme reverse transcriptase is a vital enzyme as a DNA copy is made conveniently by the help of this enzyme. Many other accurate and efficient enzymes are available, which have been manufactured by the companies and have originated from the original structure. But there accuracy is still not 100% guaranteed (Trent et al., 2005).
The PCR can be successful in totality if the flanking areas are studied and a detailed knowledge about these regions is obtained. The knowledge will enable the designing of both the backwards and the forward primers can be done specifically for the attachment. Specific reagents are required to enable successful amplification, which may be sterile water, magnesium, or other enzymes. The identification process of the pathogens can be achieved after the exponential amplification is initiated with the selection of proper reagents and adequate conditions (Erlich et al., 1991).
Phenotypic and Genotypic-based tests
The diagnostic methods are classified as either phenotypic or genotypic.
Phenotypic tests: these tests have been widely used for many years because of its cost effectiveness and availability. In comparison, this technique has problems like inconvenience in visualisation with lot of time consumed. It cannot be done by anyone but requires application by specialised scientists. Varying physiological characteristics and properties may develop due to a longer time period in the culture. Therefore differences may be observed in these pathogens as compared to those of the early virus lifecycle (Trent et al., 2005).
Genotypic tests: these tests have been further developed with the advancements in molecular biology. These techniques have been able to replace the above mentioned traditional methods. But the genotypic technique has proven to be expensive.
Another technique has now been developed since the last 20 years, which is the profiling of proteins and bacteria. This has made the characterisation of infectious agents much easy (Tang et al., 1997). .
The technique of microarrays has been created in wake of the technological advancements that have been taking place at a rapid pace. This technique has been very beneficial as it allows the analysis up to 1,000,000 oligonucleotides per cm2. . A DNA molecule is attached or spotted onto the solid surface such as glass or silicon in the microarray apparatus. The DNA spotting is executed through computer for accuracy on to the solid surface, which should not be porous at all (Trent et al., 2005).
The attached DNA spots on the glass are complementary with respect to the DNA probe, the sequence of which is attached to the solid surface. This DNA is either cDNA (complimentary DNA) or a collection of oligonucleotides.
Radioactive labels attached to the probe, are used to identify the complementary match established between the template DNA and the probe DNA. A fluorescence dye can also be used for this purpose. The technique is unique as it allows accurate and immediate results interpretation.
In the case of newly identified diseases, the microarray technology can be effectively utilised as it can be used in the comparative analysis of the patient's sample to that of the normal person. As particular genes express certain results, these certain genes can be selected for imprintation on the microarray chip. The comparative analysis of the healthy and the infected genes will render results showing differences in expressions of the genes (Trent et al., 2005).
Conclusions can be derived with the help of the knowledge on over or under expressed genes explaining the processes and the mechanism of the diseases.
This method is effectively utilised for the pathogen characterisation as it proves efficient in terms of sensitivity and specificity of the immunological procedures. The pathogens however have to display a unique property or a structure for this process to be effective which will then lead to the formation of antibody and antigen complex (Glick et al., 2003). After the formation of the complex, a spectrophotometer can be used to see the attachment to a particular substrate, thus determining the pathogen's presence.
Another technique which helps in the immunological properties is the Enzyme-Linked Immunosorbent Assay (known as ELISA). The technique is being commonly used in laboratories. The knowledge of direct attachment sites is primary as it utilizes an antibody which bonds with the antigen. The process is followed by washing to remove any unbound antigen, to which then another primary antibody is then added. This secondary antibody will fulfil two important purposes. Firstly it will bind to a particular primary antibody and secondly it will also bind with an enzyme such as the alkaline phosphatise. The enzyme will help the researchers in visually identifying the presence of a reaction in case of the binding of the secondary antibody to a secondary antibody. (Zackrisson, G et al., 1986).
A specific antibody according to the reaction as a result of the pathogen needs to be produced for the technique to work properly. The introduction of a test subject like a rabbit or a mouse will have a certain immune response, which will render the antigens specific to the antigen. Thus the antibodies can be collected through this method.
The cure developed to be effective must first address the virus and then the symptoms occurring because of the disease. The cure for the disease will then be achieved. A range of techniques such as genomics, bioinformatics, molecular biology and proteomics are available. (Warrell et al., 2005). These techniques can be used to develop a cure by keeping in mind the following factors:
Characterisation of the host target to devise ways in restricting the infectious agents resulting in harmful diseases.
The nucleotide sequencing of the pathogen.
Stop the replication in case of the pathogen being a bacterium. In case of a virus, devise ways to restrict it from attaching to the host's components.
Pathogen modifications with respect to the identification of primary sequences in the initiation and the transcription.
The disease development should be stopped by restricting the disrupted regulatory processes.