DIRECT BLOOD SMEAR

While this method is a simple screening test, it is not recommended as a regular test method. In this test, a single drop of blood is examined under the microscope for the presence of live microfilariae. If a large number of microfilariae are present, chances are at least one will be seen seen swimming by. Of course, detection of heartworm larvae in this way require large numbers of larvae for detection (less than 20-50 microfilariae per ml of blood will not be detected). More subtle infections will be missed.


DIFIL TEST AND KNOTT'S TEST

These tests represent "concentration" methods for the detection of microfilariae so that more subtle infections can be detected. In these tests, a larger amount of blood is either centrifuged or filtered to concentrate any microfilariae present. When a microfilaria test is to be done, either of these would be appropriate. Nowadays, microfilaria testing is usually done in conjunction with antigen testing.


DIPETALONEMA RECONDITUM: THE OTHER MICROFILA

Heartworm is not the only species of worm that has circulating microfilariae as a first stage larva. There is another parasite called Dipetalonema reconditum, which is transmitted by fleas, which may be picked up by the microfilaria tests. Subtle differences in the swimming characteristics, nose structure, and tail structure can be used to distinguish this harmless larva from the more serious heartworm microfilaria. Before immunotechnology, there was an element of diagnostic challenge when microfilaria were detected in the blood (was the nose the shape that would justify a harsh and possibly life-threatening treatment for heartworm infection or was it a harmless creature for which no treatment was needed?) The advent of antigen testing has made this distinction much easier to make.


ANTIGEN TESTING

Using genetic engineering, it has become possible to create extremely sensitive tests capable of detecting tiny pieces of adult heartworm skin circulating in the blood. In this way, very small numbers of adult worms can be detected, even single worm infections. This kind of testing has made it possible to detect infections in which no microfilariae are present: the so-called "Occult Infections." Most of these immunological tests are available as test kits which can be performed in the veterinarian's office while you wait.

There are many important reasons why a dog might be infected with adult heartworms yet no microfilariae can be detected:

Single sex infections or single worm infections
Younger female worms tend to be resistant to the drugs used to clear adult worms. This means that after the first treatment with anti-heartworm medication, a group of young female worms will be left. If antigen testing is not performed, one could be misled into thinking that the infection had been completely cleared.

Immunologic destruction of microfilariae
In the cat, the period in which microfilariae can be detected in the blood stream is extremely short as the immune system rapidly groups and clears these larval worms.

The dog is taking heartworm prevention medication.
Both Interceptor and Heartgard 30 are able to kill circulating microfilariae (but not adult heartworms). If for some reason, a dog is not properly tested and has missed a dose of medication, it is possible for an infection to establish yet no microfilariae will be detected. People commonly ask why they must continue annual testing in animals that are on preventive medication; this is one reason.

Since Dipetalonema microfilariae can be mistaken for heartworm larvae and since microfilariae can be transferred to unborn puppies (but adult heartworms cannot), it is felt by many specialists that no microfilaria positive dog should be treated for heartworm without a positive antigen test.


ANTIBODY TESTING

Antigen tests have limited usefulness when it comes to infections involving only a few worms. One problem is that the antigens detected by antigen tests are unique to female worms. For most tests, at least three female worms are needed for the test to show a positive result. Animals infected with only male worms will test antigen negative. To get around these limitations, antibody tests have been developed to detect the host's immune response against the parasite. This kind of breakthrough has been especially important in testing for cats in whom infection with one or two worms is the usual situation. Antibody testing may be able to detect infection sooner than can antigen testing and infections involving only male worms can be readily detected.