9. Blood group testing GERALD D. HART, MD; INGE KVAS, LCSLT; MARJA L. SOOTS, MD
Blood group SLlbstances are complex carbohydrates that may be separated from crythrocytes and may be detected after erythrocyte disintegration. ForenSIC scicntists have developed techniques for identifying the blood groups of dead persons and those of blood and tissue stains found during the course of criminal investigation. The serologic micromethod (8MM) and the inhibition agglutination test (tAT) used in such investigations are also applicable to the study of ancient materials; for example, Connolly and Harrison' used the SMM to investigate the kinship of the ancient Egyptian kings Srnenkhkare and Tutan khamen. With the 8MM, A or B antigens are adsorbed to fresh group 0 human erythrocytes; the presence of these antigens can be demonstrated by the microscopic agglutination produced by specific antisera. In previoLls stLldies we modified this technique and enhanced its sensitivity by adding an indirect Coombs' test to the original method.2 With the tAT, which detects minute amounts of blood group substances, the ability of an Llnknown material to inhibit known specific antisera is assessed. For example, if a sample of unknown blood group contains B antigen and is combined with anti-B serum Reprint requests to: Dr. Gerald D. Hart, Physician in chief, Toronto East General and Orthopaedie Hospital, 825 Coxwell Ave., Toronto, Ont. M4C 3E7
Historical notes - tuberculosis Tuberculosis, like syphilis, has had a great influence on the history of man. ihere is evidence of spinal tuberculosis in Egyptian mummies, (hid Aristotle and Hippocrates knew about the disease; thei' called it phthisis. The disease later caine to be known as consumnption. In the early 19th century it W(15 responsible for the (leath of up to a third of patients in large urban communities. The name tuberculosis caine into use after Svlvius ascribed the term tubercle to the nodular lesions found at autopsy. With the (liscol'erv of the stethoscope bi' Laennec earlier diagnosis became I)ossible, amid in 1843 Kleneke proved that tuberculosis was imifectious; he was successful iii transmitting the disease to rabbits by injecting themn with humnan C(lseoIIs material. Koch identified the organisimi in 1882 (ln(l Clemens von Pirquet imitroduced the tuberculin test in 1907.
of known potency, some of the anti-B will be bound by the antigen and will produce a measurable decrease in the potency of the remaining anti-B serum. With both the SMM and IAT caution is needed in the detection of blood group 0. If with the SMM no agglutination occurs with anti-A or anti-B serLim, the lack of reaction may be due to either the presence of group 0 sLibstance or the absence of any antigenic material. With the IAT the unknown sample is adsorbed with H antigen (a precursor blood group antigen present in highest quantity in 0 cells). Blood group 0 is suggested when there is a decrease in the anti-H potency without change in the anti-A or anti-B activity. Splenic material and dark brown sLibstance from the inside of the sigmoid sinus of Nakht (ROM I) were tested with the 8MM and IAT. Repeated testing of the splenic material with the 8MM produced no agglutination and the procedure was complicated by hemolysis of the adsorbed group 0 cells. However, when the splenic material was tested with the IAT a positive resLilt for blood group B was obtained. The sigmoid sinus material showed a positive reaction for blood group B with the serologic micromethod and with the IAT. We had no explanation for these results until we studied histologic sec-
tions of the spleen and the sigmoid sinus. The splenic material was heavily contaminated by bacterial and fungal spores that were hemolytic to the group O test cells. Hence the presence of B antigen in the splenic material could not be demonstrated by adsorption to group 0 cells, but was detectable by the JAT. Histologic sections of the sigmoid sinus showed excellent preservation of erythrocytes and absence of bacterial spores. When tested, this material gave a good source of blood group antigen and the tests were not negated by contaminating hemolytic spores. Our observation of intact erythrocytes for many centuries is not the first. Zimmerman3 reported preservation of intact erythrocytes in the pulmonary vein of a naturally preserved 2000-year-old American Indian mummy. At this time the erythrocytes isolated from Nakht are the oldest known preserved human blood cells. The histologic findings make us confident that our testing techniques are bona fide and that Nakht's blood group was B. References
A new era in the history of tuberculosis began in January 1944, when Selman A. Waxrnan of Rutgers University, together with two colleagues, announced the discovery of streptoinycin. The incidence of the disease had decreased even before sanitary laws caine into existence, however, and the death rate decreased from 400 per 100000 population in the 1830s to 80 per 100 000 ill 1925. Since Waxman's discovery the death rate in the United States has decreased to 3.1 per 100 000 population. Tuberculosis continues to be more common among the poor in areas of population congestion. Edward Livingstone Trudeau (18481915), a New York City physician, had what he took to be terminal tuberculosis and went to the Adirondack Mountains in upper New York State to spend his last few months hunting and fishing. He recovered and thus caine to believe in rest and fresh air as a form of treatment. With the help of wealthy friends and grateful patients
he established a world-famous .anitorium close to Saranac Lake, where his statue now stands; at its base are inscribed the purpose and intent of medical practice - to cure sometimes, to relieve often, but to comfort always. Norman Bethune (1890-1 939), who later found a place for himself in Chinese history, was a chest surgeon. Bethune caine to believe in the value of pneumothorax as a means of resting and curing the lung affected with tuherculosis, and he developed or modified various instruments used in general thoracic surgery and machines used to induce pneumothorax. Bethune was at one time very ill with tuberculosis and spent some time as a patient at the Trudeau sanitorium. He found that, like Thomas Mann's Hans Castorp, there was much to be learned about oneself and one's fellow mnan in the restful hours of contemnplation that seem to have been a part of the distinctlv isolated environmnent of the
476 CMA JOURNAL/SEPTEMBER 3, 1977/VOL. 117
I. CONNOLLY RC, HARRISON RD: Kinship of
Smenkhkare and Tutankhamen affirmed by serological micromethod. Nature 224: 325, 1969 2. HART GD, KYAS I, SooTs ML: Blood group
testing of skeletal remains from the North Elmham site (in press) 3. ZIMMERMAN MR: Blood cells preserved in a mummy 2000 years old. Science 180: 303, 1971
sanitoriumn.E
Blood group SLlbstances are complex carbohydrates that may be separated from crythrocytes and may be detected after erythrocyte disintegration. ForenSIC scicntists have developed techniques for identifying the blood groups of dead persons and those of blood and tissue stains found during the course of criminal investigation. The serologic micromethod (8MM) and the inhibition agglutination test (tAT) used in such investigations are also applicable to the study of ancient materials; for example, Connolly and Harrison' used the SMM to investigate the kinship of the ancient Egyptian kings Srnenkhkare and Tutan khamen. With the 8MM, A or B antigens are adsorbed to fresh group 0 human erythrocytes; the presence of these antigens can be demonstrated by the microscopic agglutination produced by specific antisera. In previoLls stLldies we modified this technique and enhanced its sensitivity by adding an indirect Coombs' test to the original method.2 With the tAT, which detects minute amounts of blood group substances, the ability of an Llnknown material to inhibit known specific antisera is assessed. For example, if a sample of unknown blood group contains B antigen and is combined with anti-B serum Reprint requests to: Dr. Gerald D. Hart, Physician in chief, Toronto East General and Orthopaedie Hospital, 825 Coxwell Ave., Toronto, Ont. M4C 3E7
Historical notes - tuberculosis Tuberculosis, like syphilis, has had a great influence on the history of man. ihere is evidence of spinal tuberculosis in Egyptian mummies, (hid Aristotle and Hippocrates knew about the disease; thei' called it phthisis. The disease later caine to be known as consumnption. In the early 19th century it W(15 responsible for the (leath of up to a third of patients in large urban communities. The name tuberculosis caine into use after Svlvius ascribed the term tubercle to the nodular lesions found at autopsy. With the (liscol'erv of the stethoscope bi' Laennec earlier diagnosis became I)ossible, amid in 1843 Kleneke proved that tuberculosis was imifectious; he was successful iii transmitting the disease to rabbits by injecting themn with humnan C(lseoIIs material. Koch identified the organisimi in 1882 (ln(l Clemens von Pirquet imitroduced the tuberculin test in 1907.
of known potency, some of the anti-B will be bound by the antigen and will produce a measurable decrease in the potency of the remaining anti-B serum. With both the SMM and IAT caution is needed in the detection of blood group 0. If with the SMM no agglutination occurs with anti-A or anti-B serLim, the lack of reaction may be due to either the presence of group 0 sLibstance or the absence of any antigenic material. With the IAT the unknown sample is adsorbed with H antigen (a precursor blood group antigen present in highest quantity in 0 cells). Blood group 0 is suggested when there is a decrease in the anti-H potency without change in the anti-A or anti-B activity. Splenic material and dark brown sLibstance from the inside of the sigmoid sinus of Nakht (ROM I) were tested with the 8MM and IAT. Repeated testing of the splenic material with the 8MM produced no agglutination and the procedure was complicated by hemolysis of the adsorbed group 0 cells. However, when the splenic material was tested with the IAT a positive resLilt for blood group B was obtained. The sigmoid sinus material showed a positive reaction for blood group B with the serologic micromethod and with the IAT. We had no explanation for these results until we studied histologic sec-
tions of the spleen and the sigmoid sinus. The splenic material was heavily contaminated by bacterial and fungal spores that were hemolytic to the group O test cells. Hence the presence of B antigen in the splenic material could not be demonstrated by adsorption to group 0 cells, but was detectable by the JAT. Histologic sections of the sigmoid sinus showed excellent preservation of erythrocytes and absence of bacterial spores. When tested, this material gave a good source of blood group antigen and the tests were not negated by contaminating hemolytic spores. Our observation of intact erythrocytes for many centuries is not the first. Zimmerman3 reported preservation of intact erythrocytes in the pulmonary vein of a naturally preserved 2000-year-old American Indian mummy. At this time the erythrocytes isolated from Nakht are the oldest known preserved human blood cells. The histologic findings make us confident that our testing techniques are bona fide and that Nakht's blood group was B. References
A new era in the history of tuberculosis began in January 1944, when Selman A. Waxrnan of Rutgers University, together with two colleagues, announced the discovery of streptoinycin. The incidence of the disease had decreased even before sanitary laws caine into existence, however, and the death rate decreased from 400 per 100000 population in the 1830s to 80 per 100 000 ill 1925. Since Waxman's discovery the death rate in the United States has decreased to 3.1 per 100 000 population. Tuberculosis continues to be more common among the poor in areas of population congestion. Edward Livingstone Trudeau (18481915), a New York City physician, had what he took to be terminal tuberculosis and went to the Adirondack Mountains in upper New York State to spend his last few months hunting and fishing. He recovered and thus caine to believe in rest and fresh air as a form of treatment. With the help of wealthy friends and grateful patients
he established a world-famous .anitorium close to Saranac Lake, where his statue now stands; at its base are inscribed the purpose and intent of medical practice - to cure sometimes, to relieve often, but to comfort always. Norman Bethune (1890-1 939), who later found a place for himself in Chinese history, was a chest surgeon. Bethune caine to believe in the value of pneumothorax as a means of resting and curing the lung affected with tuherculosis, and he developed or modified various instruments used in general thoracic surgery and machines used to induce pneumothorax. Bethune was at one time very ill with tuberculosis and spent some time as a patient at the Trudeau sanitorium. He found that, like Thomas Mann's Hans Castorp, there was much to be learned about oneself and one's fellow mnan in the restful hours of contemnplation that seem to have been a part of the distinctlv isolated environmnent of the
476 CMA JOURNAL/SEPTEMBER 3, 1977/VOL. 117
I. CONNOLLY RC, HARRISON RD: Kinship of
Smenkhkare and Tutankhamen affirmed by serological micromethod. Nature 224: 325, 1969 2. HART GD, KYAS I, SooTs ML: Blood group
testing of skeletal remains from the North Elmham site (in press) 3. ZIMMERMAN MR: Blood cells preserved in a mummy 2000 years old. Science 180: 303, 1971
sanitoriumn.E
