Francisco, Frances Lorraine R.

LABORATORY ACTIVITY 4

1. Define cancer and its clinical features and biology.

Cancer is a class of diseases in which a group of cells display uncontrolled growth (division beyond the
normal limits), invasion (intrusion on and destruction of adjacent tissues), and sometimes metastasis
(spread to other locations in the body via lymph or blood). These three malignant properties of cancers
differentiate them from benign tumors, which are self-limited, do not invade or metastasize. Most cancers
form a tumor but some, like leukemia, do not. The branch of medicine concerned with the study,
diagnosis, treatment, and prevention of cancer is oncology.

Nearly all cancers are caused by abnormalities in the genetic material of the transformed cells . These
abnormalities may be due to the effects of carcinogens, such as tobacco smoke, radiation, chemicals, or
infectious agents. Other cancer-promoting genetic abnormalities may be randomly acquired through
errors in DNA replication, or are inherited, and thus present in all cells from birth. The heritability of
cancers are usually affected by complex interactions between carcinogens and the host's genome. New
aspects of the genetics of cancer pathogenesis, such as DNA methylation, and microRNAs are
increasingly recognized as important.

Genetic abnormalities found in cancer typically affect two general classes of genes. Cancer-promoting
oncogenes are typically activated in cancer cells, giving those cells new properties, such as hyperactive
growth and division, protection against programmed cell death, loss of respect for normal tissue
boundaries, and the ability to become established in diverse tissue environments. Tumor suppressor genes
are then inactivated in cancer cells, resulting in the loss of normal functions in those cells, such as
accurate DNA replication, control over the cell cycle, orientation and adhesion within tissues, and
interaction with protective cells of the immune system.

Cancers are classified by the type of cell that resembles the tumor and, therefore, the tissue presumed to
be the origin of the tumor. These are the histology and the location, respectively. Examples of general
categories include:

 Carcinoma: Malignant tumors derived from epithelial cells. This group represents the most
common cancers, including the common forms of breast, prostate, lung and colon cancer.
 Sarcoma: Malignant tumors derived from connective tissue, or mesenchymal cells.
 Lymphoma and leukemia: Malignancies derived from hematopoietic (blood-forming) cells
 Germ cell tumor: Tumors derived from totipotent cells. In adults most often found in the testicle
and ovary; in fetuses, babies, and young children most often found on the body midline,
particularly at the tip of the tailbone; in horses most often found at the poll (base of the skull).
 Blastic tumor or blastoma: A tumor (usually malignant) which resembles an immature or
embryonic tissue. Many of these tumors are most common in children.
2. Define immunofluorescence, immunodiffusion and immunoelectrophoresis.

Immunofluorescence (IF)
is a commonly used laboratory method for detection of antigens of cells and tissues.

The fluorescent techniques are extremely specific and sensitive. This technique consists of labeling
antibody with fluorescein isothiocyantate, a fluorescent compound with an affinity for proteins to
form a complex, conjugate. The fluorescent assay includes: direct immunofluoresent assay and
indirect immunofluoresent assay.

Direct Immunofluorescent assay In this technique, Fluorescein- conjugated antibody is used to

detect antigen- antibody reactions. This method can be applied to the detection of hepatitis B virus
& chlamydia. A fluorescent microscope is required to observe the production of color; fluorescein
gives a yellow- green light.

Indirect Immunofluorescent Assay (IFA) This method is based on the fact that antibodies not only
react with homologous antigens but can act as antigens and react with antibody.

Immunodiffusion
These are of two type: single and double immuodiffusion.

Double diffusion
This technique also referred to as the Ouchterlony method, may be used to determine the relation
ship between antigen and antibodies.

Principle
Antibody dilutions and specific soluble antigens are placed in adjacent wells. If the well size and
shape, distance between wells, temperature, and incubation time are optimal, these solutions diffuse
out, bind to each other, cross-link, and form a visible precipitate at the point of equivalence
perpendicular to the axis line between the wells the precipitation bands will be compared with a
standard antigen. The precise location of the band depends on the concentration and rate of
diffusion of antigen and antibody. In a condition of antibody excess, the band will be located nearer
the antigen well. If two antigens are present in the solution that can be recognized by the antibody,
two precipitin bands form independently.

Antibodies associated with autoimmune disorders such as rheumatoid arthritis and systemic lupus
erythematosus can be identified by double diffusion.

Immunoelectrophoresis (CIE)
a variation of the classic precipitin procedure; it merely adds an electrical current to help antigens
and antibodies move to wards each other more quickly than in simple diffusion. The procedure
takes advantage of the net electric charge of the antigens and antibodies being tested in a particular
test buffer. Variables such as types of gel, amount of current, a concentration of antigen and
antibody must be carefully controlled for maximum reactivity. The sensitivity of CIE is 10 to 20
times greater than in immuno-double diffusion, however, it is more expensive than other techniques
such as immunodiffusion
3. Enumarate the different bacterial, viral, autoimmune, parasitic, and fungal diseases
and the methods of serologic detections for each.

Rheumatic fever
Glomerulonephritis
Diagnostic Tests
Culture
beta-hemolytic group A streptococci are most reliable; however, the sequalae are immunologically
mediated and do not involve actively growing bacteria.
ASO rapid latex agglutination test
Principle: Latex particles coated with Streptolysin O agglutinate when mixed with patient's serum
containing ASO antibody.

Streptozyme
Principle: Streptozyme is a passive hemagglutination assay. Newer methods use latex as the earner
particle. Immunonephelometry assays are also available.

SYPHILIS SEROLOGY
Causative Agent: Treponema pallidum subsp. pallidum, a spirochete Transmitted by direct contact
(including sexual contact) and across the placenta

Diagnosis:
1.Direct Detection
2.VDRL test, USR test, TP-PA test, FTA-ABStest

BORRELIA BURGDORFERI SEROLOGY

Causes Lynie disease, also referred to as Lyme borreliosis

Diagnosis
Culture
Serology tests- diagnosis can be made if a fourfold increase in titer is detected between an acute
serum specimen and a specimen taken 6-8 weeks later (convalescent). A more rapid method is to
detect IgM antibodies to B. burgdorferi antigens.
b. Immunofluorescence and enzyme-linked immunosorbent assays are screening methods. Positive
specimens should be confirmed by immunoblotting.
c. Immunoblot (Western blot)

Rubella (German measles)

Epstein Barr Virus

Test methods
used include latex agglutination, passive hemagglutination, ELISA, and indirect
immunofluorescence. 2. Hemagglutination inhibition test a. Rubella virus agglutinates chick RBCs.
b. Patient serum is combined with rubella antigen. If the patient has antibodies to the rubella
antigen, an antigen-antibody complex forms and it does not allow the rubella antigen to agglutinate
the indicator chick RBCs.
c. No agglutination indicates that antibodies are present.
3. Passive agglutination: Latex particles coated with rubella virus are agglutinated by rubella
antibodies, if present.
Infectious mononucleosis

VIRAL HEPATITIS SEROLOGY

Hepatitis Testing. The most widely used test method is ELISA.
HUMAN IMMUNODEFICIENCY VIRUS SEROLOGY
Laboratory Tests
ELISA tests are used to detect antibodies to HIV and HIV antigen. Repeatedly positive samples must be
confirmed by a Western blot or immunofluorescent test. 2. The Western blot assay is the confirmatory
serological test for HIV. Two of the three bands must appear for a Western blot to be considered positive:
p24, gp41,orgp!20/160. 3. Genetic probes can detect replicating viruses. 4. Reverse transcriptase—
polymerase chain reaction assays detect nucleic acid gene sequences in HIV-1 and HIV-2. 5. The indirect
immunofluorescence assay is used to detect HIV antigen in infected cells. This can also be used as a
confirmatory test.