Serology laboratory

Bacterial diagnosis

Widal test:
S.typhi and S.paratyphi A, B infect humans via the digestive tract and cause typhoid and paratyphoid fever. Antibodies
to O antigen (the body) rise in the active stage of infection and persist for few monthes. Presence of low –moderate
antibodies to H antigen (flagellar) is suggestive of expose.

Brucella direct test:

Alternate Names : Brucella Antibody Test or Titer, Brucella Serology
This is an analysis of blood to detect the presence of antibodies against Brucella (the bacteria that causes the disease
brucellosis).
IgM antibody appears soon after infection. IgG can stay for a long time.

Wright/Direct Brucella Test:

It is used to detect if there is Brucella infection or not.
Procedure:
 Put 25 μl of serum with 25 μl of reagent containing Brucella antigen
 Incubate 4 minutes at room temperature while shaking
 Read the results

Result:
If there is agglutination, this will indicate Brucella antibodies in the serum of the patient. If there was no agglutination, a
Brucella blocking test should be done to confirm the results.

Brucella blocking:
When chronic brucellosis is strongly suspected but cannot be confirmed through routine agglutination tests, an extra test
reagent (anti human globulin) may be added to the brucella antigen for additional sensitivity.

This confirmatory test is done for 2 reasons:

 Antibodies are aggregated thus blocking the antigen from reacting with them
 A patient who is taking a drug and thus the antibody titer is very low and can be detected only by using a
microscope

Procedure:

Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 Tube 6 Tube 7

NaCl 1000μl 500 μl 500 μl 500 μl 500 μl 500 μl 500 μl
 From the first tube, take 100 μl of NaCl and replace it by 100 μl of the serum to be tested
 Take 500 μl from the first tube and put it in the tube 2 and repeat the same step until the tube 6 (double
dilution). Tube 7 is considered a control tube
 Put in tubes 1 till 7, 50 μl of Brucella antigen provided by the kit
 Incubate at 56°C in water bath for one hour
 Shake and centrifuge for 10 minutes
 Wash 3 times with NaCl , shake and centrifuge for 10 minutes
 Add two drops of anti-IgG, Mix and incubate for 24 hours at 37°C
 Read the results by using a microscope

ASO / Antistreptolysin O

Also known as: ASLO Formal name: Antistreptolysin O

This test measures the amount of antibody produced in response to a recent streptococcal infection. Streptolysin O is a
toxin made by the Group A streptococcus bacteria (Streptococcus pyogenes). Antibodies made against this foreign toxin
are known as antistreptolysin O (ASO) antibodies, which appear in the blood following infection.

ASO test results can be reported in several different ways; however, the interpretation is generally the same: the higher
the result is, the more antibody that is present in the blood (unless a titer is performed, which is a ratio and therefore is
interpreted differently).
1
The ASO antibody is either absent or present in very low concentrations in patients who have not had a recent
streptococcal (strep) infection. Antibodies are produced about a week to a month after the initial strep infection. ASO
levels peak at about 4 to 6 weeks after the illness and then taper off but may remain at detectible levels for several
months after the strep infection has resolved.

Results interpretation:

If the test is negative or if ASO is present in very low concentrations, then the patient most likely has not had a recent
strep. infection. This is especially true if a sample taken 10 to 14 days later is also negative or minimal.

If the ASO level is high or is rising, then it is likely that a recent strep infection has occurred. ASO levels that are
initially high and then decline suggest that an infection has occurred and may be resolving.

Helicobacter Pylori

Also known as: H. pylori, H-pylori, H. pylori antibody test, H. pylori antigen test, H. pylori breath test
Formal name: Helicobacter Pylori

These tests are looking for evidence of an infection by a bacterium, known as Helicobacter pylori. This bacterium is
now known to be a major cause of peptic ulcer disease. H. pylori is also associated with the development of gastric
cancer.

A positive H. pylori test, antibody, antigen, or breath test signifies that you have been infected with this organism. In
recent years, scientific data support that this bacteria causes stomach ulcers and appropriate treatment can destroy the
bacteria and stop the disease.

Pronto dry test:

Pronto Dry™ is intended for use for the detection of urease enzyme in gastric mucosal biopsy specimens.

Features of the test:

 All results in one hour

 High sensitivity and specificity for accuracy
 Very cost effective
 Simple to interpret
 Easy to use
 No temperature sensitivity
 No refrigeration
 No incubation

Helicobacter pylori breath tests

H pylori diagnostic breath tests rely on a simple chemical reaction which is based on the natural behavior of the
bacteria. Naturally occurring gastric urea is made up of 99% carbon isotope 12C and 1% carbon isotope 13C. The breath
test uses urea enriched with 13C (that is, 99%).

Virtually all duodenal ulcers and 80% of gastric ulcers are associated with Helicobacter pylori infections

2
H. pylori survive in gastric acid by excreting large amounts of urease. This enzyme breaks down any urea in the
stomach to ammonia and carbon dioxide.

The ammonia neutralizes any acid found directly in the vicinity of the bacteria, allowing
the bacteria to survive.

The carbon dioxide is absorbed into the blood stream and then released from
the lungs.

Urea breath test. For the urea breath test, you swallow a capsule or drink water that
contains urea. The urea is tagged with radioactive carbon (carbon-14) or a form of carbon that is not radioactive
(carbon-13). If H. pylori bacteria are present in your stomach, they will break down the urea, eventually causing you to
exhale carbon dioxide that contains the tagged carbon. If you have no H. pylori bacteria in your stomach, the urea does
not break down and instead passes out of your body in your urine.

Syphilis

Also known as: VDRL, RPR, FTA, Darkfield microscopy

Formal name: Syphilis detection test

The test is looking for evidence of Treponema pallidum, the bacterium that causes syphilis. Syphilis is a common
sexually transmitted disease. It is easily treated but can cause severe health problems if left untreated.

The blood test detects the antibodies that the body produces to combat infection, so a positive test indicates that you
have either a current or past infection. The blood test might not find antibodies for up to three months after exposure to
the bacteria. In addition, the antibodies remain in the body for years, so if you have had a past infection with syphilis
and were treated, your test results could still be positive.

It is done using rapid test. This kit is based on the latex agglutination between antigens and antibodies.

The antibody to the syphilis bacteria (reagin) is not present. This is called a
Normal nonreactive or negative result.
The antibody reagin is present. This is called a reactive or positive test.

3
 Abnormal

TORCH diagnosis

Toxoplasmosis

Antibody Detection
The detection of Toxoplasma-specific antibodies is the primary diagnostic method to determine infection with
Toxoplasma. Toxoplasma antibody detection tests are performed by a large number of laboratories with commercially
available kits.An algorithm for the immunodiagnosis of toxoplasmosis for individuals greater than one year of age is
shown in the following figure. The IFA and EIA tests for IgG and IgM antibodies are the tests most commonly used
today. Persons should be initially tested for the presence of Toxoplasma-specific IgG antibodies to determine their
immune status. A positive IgG titer indicates infection with the organism at some time. If more precise knowledge of
the time of infection is necessary, then an IgG positive person should have an IgM test performed by a procedure with
minimal nonspecific reactions, such as IgM-capture EIA. A negative IgM test essentially excludes recent infection, but
a positive IgM test is difficult to interpret because Toxoplasma-specific IgM antibodies may be detected by EIA for as
long as 18 months after acute acquired infection.

Rubella

Also known as: German measles, 3-day measles

Formal name: Rubella antibodies, IgM and IgG

This test measures the presence of rubella antibodies in the blood. Rubella antibodies are produced in response to an
infection by the rubella virus. There are two types of rubella antibodies: IgM (short-term) and IgG (long-term). The
first type to appear in the blood after exposure is the IgM rubella antibody. The level of this protein in the blood rises
and peaks within about 7 to 10 days after infection and then tapers off over the next few weeks, except in an infected
newborn where it may be detected for several months to a year. The IgG rubella antibody takes a bit longer to appear
than the IgM, but once it does, it stays in the bloodstream for life, providing protection against re-infection.

CMV / Cytomegalovirus

Formal name: Cytomegalovirus

Cytomegalovirus (CMV) testing is used to determine whether someone is currently, or has recently been, infected with
CMV. Testing involves either a measurement of CMV antibodies (specific proteins created by the body’s immune
system in response to the virus) or the detection of the virus itself, through culturing and growing the virus or by
amplifying the genetic material from the virus itself.

Antibody testing:

There are two types of CMV antibodies found in the blood: IgM and IgG. IgM antibodies are the first to be produced by
the body in response to a CMV infection. They are present in most individuals within a week or two after the initial
exposure. IgM antibody production rises for a short time period and then tapers off.

Levels of IgG rise during the active infection, then stabilize as the CMV infection resolves and the virus becomes
inactive. Once a person has been exposed to CMV, they will have some measurable amount of CMV IgG antibody in
their blood for the rest of their life. CMV IgG antibody testing can be used, along with IgM testing, to help confirm the
presence of a recent or previous CMV infection.
4
 Herpes

Also known as: Genital Herpes, Oral Herpes, Cold Sores

Formal name: Herpes Simplex Virus, Type1 and Type 2 (HSV-1 and HSV-2)

Herpes simplex testing is performed to identify an acute herpes infection or to detect herpes antibodies, an indication of
a previous exposure to herpes. One of the most common viral infections, herpes simplex virus (HSV) exists in two
types, HSV-1 and HSV-2. Both types are contagious and periodically cause small fever blisters (vesicles) that break to
form open lesions. HSV-1 primarily causes blisters or “cold sores” around the oral cavity and mouth, while HSV-2
usually causes lesions around the genital area.

HSV antibody testing. Antibodies to HSV are specific proteins that the body creates and releases into the bloodstream
to fight the infection. HSV IgM antibody production begins several days after a primary (initial) HSV infection and may
be detectable in the blood for several weeks. HSV IgG antibody production begins after HSV IgM production.
Concentrations rise for several weeks, fall, and then stabilize in the blood. Once someone has been infected with HSV,
they will continue to produce small quantities of HSV IgG. HSV antibody testing can detect both viral types (HSV-1
and HSV-2) and tests are available that can detect the early IgM antibodies as well as the IgG antibodies that remain
forever in those who have been exposed.