METHODS FOR THE DETERMINATION OF PNEUMOCOCCUS TYPES

Abstract

The determination of pneumococcus types in lobar pneumonia is of value in the field of prognosis and as a prerequisite for specific serum therapy. The method for the determination of types should be as rapid as possible and a standard technique should be employed. The most satisfactory method is by the intraperitoneal inoculation of a mouse with the patient's sputum, by which means a rapid and abundant growth of the pneumococcus is obtained and secondary organisms are rapidly eliminated in most instances. The diagnosis of type is made directly on the peritoneal exudate. Certain factors in the method commonly used have interfered with the rapid determination of types in an appreciable number of cases, notably the growth of other organisms in the peritoneal exudate together with the pneumococcus, and some confusion has arisen because occasional strains of pneumococci have been encountered that show cross agglutination reactions when undiluted immune serum is used. Such reactions have been shown to be due to a limited zone of non-specific immunity and they in no way invalidate the classification of the pneumococci into sharply defined immunological groups. The optimum dilutions of serum have been determined that will agglutinate all type strains of pneumococci and fail to cause any cross agglutination reactions when mixed with equal amounts of pneumococcus cultures and incubated for I hour at 37°C. They are a 1:20 dilution of Serum I, a 1:20 dilution of Seruifa II, and a 1:5 dilution of Serum III. For the diagnosis of Subgroup II pneumococci undiluted Type II serum is required. To obviate the other difficulties of the method commonly used a new method for the determination of types has been devised. It depends upon the fact that there is produced by the growth of the pneumococcus a soluble substance which is present in the peritoneal exudate of the mouse in sufficient quantity to give a specific precipitin reaction with the homologous immune serum. The precipitin method can be used in all instances in which the determination of types by the agglutination method is possible, and it possesses certain distinct advantages which make it available when the agglutination method is impracticable. It is of particular value as a time-saving device in those instances where the presence of other organisms together with the pneumococcus in the peritoneal exudate causes a delay of 18 hours or more before the type of pneumococcus can be definitely established. It is therefore recommended as the method of choice in all cases. If desired, both the agglutination and precipitin methods may be applied to the same specimen of peritoneal washings.