By John G. Heemstra, M.A., and Willis F. Stanage, M.D.
Laboratory and Pediatric Sections, Yankton Clinic, Yankton, South Dakota
ADDRESS: (J.G.H.) 400 Park Avenue, Yankton, South Dakota.
753
PEDIATRICS, April 1959
A
FINGER-TIP,
MICRO
HETEROPHILE
AGGLUTINATION
TECHNIC
T
HE DIFFICULTY and trauma in obtainingvenous blood from many pediatric pa-tients often precludes laboratory proce-dunes which may be valuable diagnostic aids. The heterophile agglutination test is omitted many times for this reason. An
ac-curate and simple means of determining the
hetenophile titer with finger-tip blood will allow many children to be followed more closely by the laboratory than would be possible using venous blood.
Various modifications have been made in an attempt to simplify the procedure. Rap-paport and Skariton1 and Glassy2 reported methods which decrease the reading time.
A qualitative
and
semiquantitative
method
which decreased the volume of blood re-quired was described by Evans.3 A method for the preservation of sheep cells was de-scnibed by Cox,4 which eliminated the washing of sheep cells prior to use. In this paper, a finger-tip method is presented which is simple and has the same range of accuracy as the regular heterophile
agglu-tination test.
MATERIAL AND METHODS
Fresh physiologic saline solution is used as the diluent throughout the entire test. Sheep cells preserved in the usual manner are freshly washed three times and used in a 2% saline suspension.
Finger-tip blood is collected in a wax de-pression cup and 0.1 ml of whole blood is
measured into a test tube (13 mm by 100 mm)
containing 0.9 ml of saline solution. Just prior
to use, a 0.1 ml-Kimax pipette is flushed with saline solution to retard clotting of blood in the pipette. The diluted blood is centrifuged to remove the cells and the supernatant serum-saline mixture inactivated at 56#{176}Cfor 30 mm-utes.
To a series of 10 Kahn tubes, 0.25 ml of saline solution is added to each except the first tube. To the first and second tubes, 0.25 ml of inactivated serum-saline is added. Twofold serial dilutions are made from the second tube to the last tube. To each tube, 0.1 ml of washed sheep cells is added and the tubes shaken thoroughly. They are then incubated at room temperature for 2 hours and the degree of
agglutination read.
Determination of Titer
The standard method for determining the hetenophile agglutination titer depends upon the final dilution of serum after all reagents are added. Normally, the first tube contains 0.05 ml of serum, 0.2 ml of saline solution and 0.1 ml of sheep cell suspension. This is 0.05 ml of serum in a total volume of 0.35 ml or the equivalent of a 1 to 7 dilution. Twofold dilution of serum in the tubes produces
dilu-tions of 1:14, 1:28, 1:56, 1:112, 1:224, 1:896
and 1:1,792.
Using the finger-tip modification, 0.1 ml of whole blood is added to 0.9 ml of saline solu-tion. The cell volume is assumed to be 50%;
therefore, the serum volume is 0.05 ml in a total volume of 1 ml or a dilution of 1 :20. With the addition of 0.1 ml of sheep cells to a total volume of 0.25 ml of serum-saline mixture, a dilution of 1 to 28 is achieved for the first tube. The following dilutions are 1 :56, 1:112, 1 :224, 1 :448, 1 :896 and 1:1,792.
EXPERIMENTAL
RESULTS
TABLE I
A COMPARISON OF FINGER-TIP AND REGULAR HEr-EROPHILE AGGLUTINATION METHODS, SHOWING
EQUAL TITERS
TABLE III
A COMPARISON OF FINGER-TIP AND REGULAR HETERO.
PHILE AGGLUTINATION METHODS, SHOWING
FINGER-TIP TITER ONE TUBE HIGHER
754 HETEROPHILE AGGLUTINATION
No. of Patients
Heterophile Aggl utinalion Titer
Finger-tip Regular
.51 <28w <28
21 28 28
19 56 56
18 112 112
10 224 224
1 448 448
aS 896 896
6 1,792 1,792
7 3,584 3,584
Total 138
* The reciprocal of the highest dilution showing ag-glutination.
DISCUSSION
One hundred eighty-one duplicate het-erophile agglutination titers were done, com-paring the finger-tip micro technic and the regular venous method; 138 resulted in the
same titer. There were 43 which showed
a difference in titer of one dilution. In one,
not recorded in the tables, there was a
dif-ference of more than one dilution. This rep-resented a patient with a finger-tip titer of 1 :896 and a regular titer of 1:224.
Three patients, or 1.6%, showed a
finger-TABLE II
A COMPARISON OF FINGER-TIP AND REGULAR
HETERO-PHILE AGGLUTINATION METHODS, SHOWING
FINGER-TIP TITER ONE TUBE LOWER
f Patients
Iteterophile Aggi ntination Titer
No. o
Finger-tip Regular
3 28 56
2 56 112
.5 112 224
No. f Patients
Helerophile Aggl utinntion Tiler o
Finger-lip Regular
5 56 28
2 112 .56
4 224 112
2 448 224
2 896 448
1 1,792 896
Total 16
tip titer of less than 1 :56 when the regular titer was 1 :56. All the other finger-tip ag-glutinations, in the significant range, were comparable to the regular heterophile
ag-glutination titer. As differences of one
dilu-tion are not significant in this serologic tech-nic, the finger-tip micro heterophile technic is a favorable and accurate method to use in pediatric practice.
As an established policy all finger-tip micro heterophile titers of 1 : 112 or higher are repeated. Venous blood is obtained for the repeat test and the titer is rechecked
using the regular heterophile agglutination technic. Differential absorption studies are done on this specimen at this time.
SUMMARY
A finger-tip micro heterophile agglutina-tion technic is described. Figures showing the comparison between the finger-tip tech-nic and the regular heterophile technic are given. The finger-tip micro heterophile titer compares favorably with the regular tech-nic and is suitable for use with pediatric
pa-tients and other individuals from whom it
SPECIAL ARTICLES 755
CARL H. SMITH, M.D.
heterophile antibody slide test. U. S. Armed Forces M.
J.,
3:947, 1952.3. Evans, A. S. : A simplified “quantitative”
method for heterophile antibody
deter-mination using capillary blood and a white cell pipette.
J.
Lab. & Clin. Med.,32:1278, 1947.
4. Cox, C. D. : Preservation of sheep
erythro-cytes and their use in a rapid plate titra-tion of heterophile antibodies in infectious
mononucleosis.
J.
Lab. & Clin. Med., 48: 298, 1956.LEUKAMIE IM KINDESALTER. BEITRAGE ZUB
MORPHOLOGIE, KLINIIC PATHoPmsIoLocn
UND Tiinipni,
J.
Oehme, M.D., W. Jans-sen, M.D., and Ch. Hagitte, M.D. Leipzig, Veb Geong Thieme, 1958, 170 pp.Based on an experience of 191 cases of
leukemia in the past 13 years, the authors have
prepared a well-organized book dealing with the essential features of leukemia in childhood.
As would be expected the bulk of cases are the
acute leukemias which they term acute
leuko-cytic, undifferentiated or paraleukoblastosis; a
small number are acute promyelocytic, chronic myelocytic and rare types. In common with the experience from other clinics the incidence
of leukemia rose from 0.159% in 1946 to 0.546%
in 1956.
The authors have written a readable and
timely text dealing concisely with the following
subjects : pathologic anatomy and hematology,
clinical aspects, individual organ changes,
leu-kemia and infection, differential diagnosis, the
newer knowledge relating to etiology and
therapy, metabolic aspects, prognosis and
psy-chologic aspects in management. In each
chap-ter current information has been assembled, this being especially noticeable in discussion of
etiology and therapy. Endogenous and
exoge-nous pathogenetic factors are discussed
includ-ing genetic and immunologic aspects, effects of
radiation and the most recent experimental observations on a viral etiology. Individual clinical features are well delineated such as the
differential diagnosis between rheumatic fever
and leukemia and leukemoid reaction versus
leukemia. The reviewer was particularly
im-pressed with the psychologic management,
es-pecially the emotional factors in the
relation-ship of patients, physician, attendants, parents and relatives.
The book is replete with beautiful color
plates with identification of individual cells,
roentgenograms and photographs of gross and
microscopic pathology of tissues and organs.
