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Antisperm Antibody Test

Antisperm Antibody Test

An antisperm antibody test looks for special proteins (antibodies) that fight against a man’s sperm in blood, vaginal fluids, or semen. The test uses a sample of sperm and adds a substance that binds only to affected sperm.

Semen can cause an immune system response in either the man’s or woman’s body. The antibodies can damage or kill sperm. If a high number of sperm antibodies come into contact with a man’s sperm, it may be hard for the sperm to fertilize an egg. The couple has a hard time becoming pregnant. This is called immunologic infertility.

A man can make sperm antibodies when his sperm come into contact with his immune system. The testicles normally keep the sperm away from the rest of the body and the immune system.

A woman can have an allergic reaction to her partner’s semen and make sperm antibodies. This kind of immune response is not fully understood but may affect fertility. This is a rare cause of infertility.

– The antisperm antibody test may be done if:
A cause for infertility cannot be found. Experts disagree about the usefulness of the test because the result may not change the treatment.
The results from another fertility test are not clear.

Sample :

For women, a blood sample is taken from a vein in the arm.

For men, a semen sample is collected after the blood and vaginal fluid samples are taken. You should not release your sperm (ejaculate) for 2 days before the test. It is important to not go longer than 5 days before the test without ejaculating.

Results :

An antisperm antibody test looks for special proteins (antibodies) that fight against a man’s sperm in blood, vaginal fluids, or semen. The higher the level of antibody-affected sperm found in the semen, the lower the chance of the sperm fertilizing an egg.

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Cyclic Citrullinated Peptide Antibody (Anti-CCP)

Cyclic Citrullinated Peptide Antibody (Anti-CCP)

Cyclic citrullinated peptide antibodies are autoantibodies produced by the immune system that are directed against cyclic citrullinated peptides (CCP). This test detects and measures anti-CCP antibodies in the blood.

Citrulline is naturally produced in the body as part of the metabolism of the amino acid arginine. However, in joints withrheumatoid arthritis (RA), this conversion may occur at a higher rate. Citrulline changes the protein structure and can trigger an immune response, producing autoantibodies against joint proteins. The CCP antibody test helps to diagnose RA and can be useful in identifying people with a more rapidly erosive form of the disease.

RA is a chronic, systemic autoimmune disease that causes inflammation, pain, stiffness, and destructive changes in the hands, feet, and other joints throughout the body. It can affect anyone at any age, but it usually develops between the ages of 40 and 60, and about 75% of those affected are women. The course of RA and its prognosis are variable. It may develop and progress slowly or rapidly. It may go into remission in some people and, in a few, it may go away. Left untreated, RA can shorten a person’s lifespan and can, within a few years, leave many of those affected too disabled to work.

A cyclic citrullinated peptide (CCP) antibody test may be ordered along with or following a rheumatoid factor (RF) test to help diagnose rheumatoid arthritis (RA) and to assess the severity and probable course of the disease (prognosis). Inflammatory markers may also be measured at this time, such as ESR and C-reactive protein (CRP).

A CCP antibody test is primarily ordered along with an RF test when someone has signs and symptoms that may be due to previously undiagnosed inflammatory arthritis or has been diagnosed with undifferentiated arthritis. It may be ordered as a follow-up test to a negative RF test when clinical signs and symptoms lead a health practitioner to suspect RA. RA usually affects multiple joints symmetrically. Signs and symptoms may include:

• Painful, warm, swollen joints of the hands and wrists most commonly
• Pain sometimes affecting elbows, neck, shoulders, hips, knees, and/or feet
• Stiffness of affected joints in the morning that improves during the course of the day
• Fatigue
• Fever
• Development of nodules under the skin, especially at the elbows.

sample :

whole blood.

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Antistreptolysin O Titer ( ASOT ) Test

Antistreptolysin O Titer ( ASOT ) Test

Antistreptolysin O (ASO) is an antibody targeted against streptolysin O, a toxic enzyme produced by group A Streptococcusbacteria. ASO and anti-DNase B are the most common of several antibodies that are produced by the body’s immune system in response to a strep infection with group A Streptococcus. This test measures the amount of ASO in the blood.

The ASO test is primarily used to help determine whether a recent strep infection with group A Streptococcus:

• Is the cause of a person’s or glomerulonephritis, a form of kidney disease
• Caused rheumatic fever in a person with signs and symptoms

The test may be ordered by itself or along with an anti-DNase B, another test used to detect recent strep infections.

In most cases, strep infections are identified and treated with antibiotics and the infections resolve. In cases where they do not cause identifiable symptoms and/or go untreated, however, complications (sequelae), namely rheumatic fever and glomerulonephritis, can develop in some people, especially young children. The test, therefore, is ordered if a person presents with symptoms suggesting rheumatic fever or glomerulonephritis and has had a recent history of sore throat or a confirmed streptococcal infection.

Some symptoms of rheumatic fever may include:

• Fever
• Joint swelling and pain in more than one joint, especially in the ankles, knees, elbows and wrists, sometimes moving from one joint to another
• Small, painless nodules under the skin
• Rapid, jerky movements (Sydenham’s chorea)
• Skin rash
• Sometimes the heart can become inflamed (carditis); this may not produce any symptoms but also may lead to shortness of breath, heart palpitations, or chest pain.

sample :

Whole blood.

-ASO antibodies are produced about a week to a month after an initial strep infection. The amount of ASO antibody (titer) peaks at about 3 to 5 weeks after the illness and then tapers off but may remain detectable for several months after the strep infection has resolved.

-An elevated titer of antibody (positive ASO) or an ASO titer that is rising means that it is likely that the person tested has had a recent strep infection. ASO titers that are initially high and then decline suggest that an infection has occurred and may be resolving.

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METHODS OF CULTIVATION OF VIRUSES

METHODS OF CULTIVATION OF VIRUSES

Since viruses are obligate intracellular parasites they have to be grown in living cells. There are three systems for their cultivation:

• Tissue culture.
• Embryonated eggs.
• Intact animals.

1– Tissue Culture

Pieces of animal or human tissues are trypsinized to get separate cells. These are grown in presence of growth medium containing serum, on glass or plastic tubes, bottles or plates with a flat side. Amono layer or sheet of cells is formed on the flat side of the contains within few days.

Viruses are inoculated on the monolayer. There are three types of tissue culture.

• Primary cell lines: These are prepared from organ fragments e.g. monkey kidney. Such cells can only divide for several passages ( 4-6 ) and then degenerate.
• Human diploid cell lines: These are usually fibroblasts derived from human embryo tissues. They have diploid number of chromosomes. They grow rapidly and can be sub cultured up to about 50 passages in culture e.g. human embryo lung tissues.
• Continuous cell lines: These are derived from tumbler cells and they can divide indefinitely e.g. Hela cells which are derived from carcinoma of the cervix.

Detection of virus replication in cell culture

1- Cytopathogenic effects (CPE): These are changes in cells that can be observed microscopically:

a- cell death and detachment from the glass surface is produced by many viruses e.g. poliovirus.

b- Rounding and grape – like cluster formation is produced by adeno virus.

c- Syncytium or multinucleate giant cell formation are characteristic of measles or mumps.

d- Cell transformation: The cell lose the property of contact inhibition present in normal cells and pile up to form foci of malignantly transformed cells e.g. when infected with tumor viruses.

2- Plaque formation: Plaques are virally infected areas in tissue culture monolayer. They can be seen by the naked eye as unstained areas when using vital stains.

3– Inclusion bodies: These are intranuclear or intracytoplasmic structures or intracytoplasmic structures.

Which may appear in virus infected cells and can be seen by the light microscope. They are often the site of virus replication. There presence is of diagnostic value e.g. the ” Negri bodies ” in nerve cells of rabid animals.

4- Haem absorption: when RBCs are added to infected cells they will appear as losettes or clumps an the areas where the virus is growing. This is useful in haemagglutinating viruses e.g. influenze virus.

5- Fluorecent – antibody staining: Infected cell sheets on cover slips or microtiter plates may be treated with fluorescien labeled specific antibody and examined for positive fluorescence.

6– Inter ference: In some viruses which do not produce CPE their growth can be proved by their ability to interfere with the growth of mother CPE producing virus.

7- Detection of viral antigens by serology: soluble antigens which diffuse in the nutrient medium or those released after freeing and thawing of tissue culture cells, can be detected by any serologic method including complement fixation, haemagglutination… etc.

8- Neutralization tests: Neutralization of the effects of virus on tissue culture by specific antisera can be used to identify and type the virus isolated.

2- Embryonated Eggs

Different viruses can grow in various cavities of Embryonated eggs or in the developing embryo it self. The age of the embryo used and the site of inoculation vary according to the virus inoculated.

Choriallantoic membrance inoculation is used in pox and herpes viruses. The influenza virus can readily grow in the amniotic sec and in the respiratory cells of the embryo.

3 – Intact Animal

 Animal inoculation was mainly used in the past when tissue culture methods were not known. However, animal inoculation is still used for studying viral oncogenesis, pathogenesis of viral diseases, immune response to viruses and for primary isolation of some viruses. The white sucking mouse is the most widely used, it is susceptible to encephotalitis viruses by the intracerbral route.

METHODS OF CULTIVATION OF VIRUSES

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HEPATITIS (D,E,G)

HEPATITIS (D,E,G)

-HEPATITIS D VIRUS ( HDV )

( Delta hepatitis ) :

HDV is a defective RNA virus that replicates only in cells also infected by HBV, because HDV uses the surface antigens of HBV (HBsAg) as its envelope. HBV provides a rescue function for HDV. Its mode of transmission is similar to HBV. The incubation period is 2-112 weeks Hepatitis in chronic HBV carriers who become super infected with HDV is more severe and the incidence of complications e.g. chronic active or fulminating hepatitis is much higher. Super infection or confection with the two viruses occurs mainly in those receiving repeated blood transfusions pooled plasma as well as drug addicts.

* Diagnosis: is done by detection of anti HDV antibodies or antigens. PCR for detection of viral RNA.

* Prevention of HBV infection by vaccination will consequently prevent HDV infection.

-HEPATITIS E VIRUS ( HEV ) :

HEV is anon enveloped single stranded RNA virus tentatively classified as a member of the calciviry family.

It infects by the faeco – oral route. Water – borne epidemics are reported. It resembles hepatitis “A” clinically with the exception that it causes high mortality rate in pregnant women.

Diagnosis: by detection of HEV specific IgM or arising titer of IgG by RIA or ELISA

-HEPATITIS G virus ( HGV ) :

HGV or GB type C virus was isolated in 1996 from patient with post transfusion hepatitis. It is a member of the flavivroy family, as is HCV.

Unlike HCV, its role as a cause of hepatitis or any other disease is still undecided. It is instead, well established that the virus is wide spread in human and can persist and circulate in the blood of infected persons in the absence of any signs or symptoms.

It is diagnosed by detecting viral RNA by PCR, or viral antibodies.

TTV and SEN virus are newly discovered viruses in the blood  of non A to G post transfusion hepatitis patients ( 1997 and 1999 respectively ). The viruses are being characterized and their clinical significance remains to be determined.

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HEPATITIS C VIRUS ( HCV )

It s a member of flavi virus family. It is a single stranded RNA enveloped virus. There are many serotypes.

* Transmission: is mainly parenteral as in HBV. Sexual and prenatal transmission has been very difficult to document and considerably less efficient than for HBV. However, it was reported possible with severely immune suppressed HIV patient.

There is no evidence of insect vector. In 10 – 50 % of cases the source of HCV infection is not known.

* Epidemiology: HCV infection is usually mild or a symptomatic. However, chronic carriage rate is higher than in HBV and 75% of cases progress to chronic disease i.e. chronic active hepatitis, cirrhosis and HCC.

* Laboratory diagnosis:

1- Detection of anti HCV antibodies in blood by ELISA and confirmation by a more specific test RIBA

( Recombinant Immune blot assay ).

2- Detection of viral RNA by PCR which indicates the presence of circulating virus i.e. active infection. It is useful in monitoring effect of anti viral therapy by measuring the viral load.

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HEPATITIS B VIRUS (HBV)

HEPATITIS B VIRUS (HBV)

HBV causes serum hepatitis. It is a DNA virus classified as a “Hepadna” virus. There morphologic forms are known.

1- 22nm spherical particles. These are the commonest.

2- 22nm tubular particles that are over 200 nm long.

The two are made up of hepatitis B surface antigen (HB_sAg)

3- A large 42 nm spherical pratical ( dane partical).

These are the complete virions of HBV and consist of an outer envelope (HBsAg) which surrounds an inner core of nucleocapsid protein( HBcAg) which surrounded the viral DNA and DNA Polymerase. (HBeAg) is also apart of the inner core.

• mode of transmission :
  1. parenteral by blood and plasma transfusion   or by the prick of contaminated needles. Risk groups include, drug abusers, medical  personnel , renal dialysis patients and those receiving transfusion repeatedly.
  2. sexual contact as the virus is found in semen and vaginal secretion.
  3. perinatal transmission from mothers with hepatitis B to the new born during birth or breast feeding

*pathogenesis :

After enteing the blood , the virus infects hepatocytes causing necrosis and inflammation. Immune attack against viral antigens an infected  hepatocytes is  mediated by cytotoxic T cell casing hepatocellular cell mediated tissue injury .this is most probably the main cause of damage , since HBV does not cause acytopathic effect. The chronic sequelae are most probably due to autoimmune reactions to liver membrane antigens induced by the virus.

• clinical findings  and outcome of infection :

many of HBV infection  are asymptomatic and are detected only by the presence of antibodies to HBsAg. After exposure there is along incubation period 2-6 months which is much longer than that of Hepatitis A. The symptoms are more severe in hepatitis B than in HAV. And can lead to life threatening complication. Unlike hepatitis A. about 10% of hepatitis B patient become chronic carriers with persistent HBV antigenaemia, which is due to persistent infection of hepatocytes.

The virus is present in the blood and other body fluids e.g. semen, saliva, vaginal secretions and milk. Most carriers active hepatitis , which can lead to cirrhosis, fulminate hepatitis and death.

A high rate of hepatocellular carcinoma (HCC) ,occurs in chronic carriers. During virus replication, it is found that some copies of viral DNA are intergratd into cell DNA which may explain the malignant predisposition  of such infection. recovery is associated with life long immunity mediated by antibodies to HBsAg.

*laboratory diagnosis :

1- increased level of liver enzymes.

2-detection of hepatitis markers i.e . antigens and antibodies in blood by ELISA.

* Antigens :

– HBsAg : is detected during the incubation period and during active disease ,it may remain for months  or years in carriers.

–HBcAg : is not detectable in serum. But can be found in the nuclei of liver cells .

–HBeAg : is present during the I.P., in the acute stage of the disease and in some chronic carriers. Its presence indicates that the serum contains high concentration of infective HBV and is highly contagious.

* Antibodies:

– HBs Ab: appears after disappearance of HBsAg and its presence indicates immunity.

– HBc Ab :appears with the onset of clinical disease and remains after recovery. It is also present in chronic disease.

-Anti – HBc IgM: detection confirms the diagnosis of recent infection.

There is a period of several weeks when HBs Ag has disappeared but HBsAb is not yet detectable. This is the window phase.

At this time, the HBcAb is always positive and can be used to make the diagnosis.

– HBe Ab: appears as HBe Ag disappears and it indicates recovery.

3- Detection of viral DNA by PCR.

4- Detection of viral DNA polymerase.

* Prophylaxis:

Pre – exposure prophylaxis by active vaccination of those at high risk. It is recommended for all newborns as part of their immunization schedule.

• plasma derived vaccine ( Hepta vax B ): This is prepares by purifying HBsAg from pooled plasma from healthy HBs Ag positive carries. The virus is inactivated. It is safe and effective in preventing HBV infection.
• Recombinant hepatitis B vaccine ( Recombivax ) is amore recent vaccine and has replaced the Heptavax B in several parts of the world. It contains HBsAg produced in yeast cells by the recombinant DNA technique. The vaccine is given in 3 intramuscular injections in the deltoid region at 1,2 and 6 months.

* Post – exposure prophylaxis: persons exposed by prick of contaminated needle or other wise or infants born to HBsAg positive mothers should immediately receive both hepatitis B specific immunoglobulin ( HBIG ) and hepatitis B vaccine, given simultaneously at different sites.

Other control measures include:

• proper choice of blood donor i.e. free of HBsAg.
• Use of plastic disposable syringes.

Treatment:  Alpha interferon is used in treatment of chronic hepatitis B.

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HEPATITIS A VIRUS (HAV)

HEPATITIS A VIRUS (HAV)

HAV was classified as an enterovirus type 72. It is now placed in anew genus in the picornavirus family called “Hepatovirus”. It is an RNA, icosahedral virus 27-30nm in diameter. Only one serotype is known. The virus is relatively heat resistant, it with stands 60c⁰ for 1 hour It is destroyed by boiling for 5 minutes. It is pathogenic to primates e.g. man and chimpanzees. It grows on primary cell lines of primate liver and human diploid cells

• Epidemiology: It is transmitted by the faecal- oral route. Incubation period is 2-6 week i.e. shorter than that of HBV. The manifestations are the same in all hepatitis viruses i.e. jaundice, fever, abdominal pain and anorexia HAV infection is usually milder than HBV many cases are non–icteric or asymptomatic. Recovery occurs without complications; no carrier, no chronic hepatitis or HCC and is followed by long lasting immunity.

The virus is found in the stools 2 weeks before and 2 weeks after the jaundice appears.

It affects mainly children and youg adults (5-15 years)

It may occur in sporadic or epidemic forms in summer comps or schools. It occurs mainly in the autumn

• laboratory diagnosis

1. Increased level of liver enzymes
2. Detection of HAV IGM or arising titre of IgG by RIA or ELISA is diagnostic of recast infection
3. PCR is applicable, but is not routinely used.
4. Detection of the virus in stools by immunoelectron microscopy is not routinely used.

• Prophylaxis:

Inactivated vaccine “Havrix” is licensed for use since 1995. It is recommended for children over over 2 years and is given intra muscularly in the deltoid region in 3 doses at 0.1 and 6 month the vaccine may be given to travelers, Armed forces and medical staff. Gamma globulin if given early after exposure renders the infection milder or sub clinical.

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Antisperm Antibody Test

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Antisperm Antibody Test

Antisperm Antibody Test

An antisperm antibody test looks for special proteins (antibodies) that fight against a man’s sperm in blood, vaginal fluids, or semen. The test uses a sample of sperm and adds a substance that binds

 only to affected sperm.

Semen can cause an immune system response in either the man’s or woman’s body. The antibodies 

can damage or kill sperm.

 If a high number of sperm antibodies come into contact with a man’s sperm, 

it may be hard for the sperm to fertilize an egg. The couple has a hard time becoming pregnant. 

This is called immunologic infertility.

A man can make sperm antibodies when his sperm 
come into contact with his immune system.

 The testicles normally keep the sperm away from the rest
 of the body and the immune system.

A woman can have an allergic reaction to her partner’s semen and make sperm antibodies. 

This kind of immune response is not fully understood but
 may affect fertility. 

This is a rare cause of infertility.

– The antisperm antibody test may be done if:

A cause for infertility cannot be found. Experts disagree about
 the usefulness of the test

 because the result may not change the treatment.

The results from another fertility test are not clear.

Sample :

For women, a blood sample is taken from a vein in the arm.

For men, a semen sample is collected after the blood and vaginal fluid samples are taken.

 You should not release your sperm (ejaculate) for 2 days before the test.

 It is important to not go longer than 5 days before the test without ejaculating.

Results :

An antisperm antibody test looks for special proteins (antibodies) that fight

 against a man’s sperm in blood, vaginal fluids, or semen. 

The higher the level of antibody-affected sperm found in the semen, the lower the chance of the

 sperm fertilizing an egg.

                  Antisperm Antibody Test

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