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10 (3-7%) of the 268 transfused patients were seropositive forHTLV-I. No serum was seropositive for HIV-1. Contemporaryrecords were available for 2 of these patients. Patient A (female,born 1901) had positive sera collected in July and November, 1969.Patient B (male, born 1927) had positive sera collected in October,1969, and in January and June, 1970. The discharge diagnoses inthese two patients were very complex but neither had clinicalconditions thought to be associated with HTLV-I. Case B had ahistory of post-transfusion hepatitis; hepatitis B was common intransfused Philadelphia patients in 1969.

Reports from the 1980s suggest that in transfused patients in theUS HTLV-I antibodies were found in 30% or SO.2,31t seems thatHTLV-I was present in transfused patients in the Philadelphia areaas early as 1969, and the frequency (37%) has not changed much.This makes it unlikely that the virus has been imported recently orhas increased in prevalence due to recent changes in behaviour or inmechanisms of transmission.

Supported by USPHS grants CA-40737, RR-05895, and CA-06927 fromthe National Institutes of Health and by appropriation from Commonwealthof Pennsylvania.

Fox Chase Cancer Center,Philadelphia, Pennsylvania 19111, USA

B. S. BLUMBERGS. H. BLANTOND. I. SHRAGER

National Cancer Institute,Bethesda, Maryland W. A. BLATTNER

1. Williams AE, Fang CT, Slamon DJ, et al. Seroprevalence and epidemiologicalcorrelates of HTLV-I infection in US blood donors. Science 1988; 240: 643-46.

2. Jason JM, McDougal JS, Cabradilla C, Kalyanaraman VS, Evatt BL. Human T-cellleukemia virus (HTLV-I) p24 antibody in New York City blood productrecipients. Am J Hematol 1985; 20: 129-37.

3. Minamoto GY, Gold JWM, Scheinberg DA, et al. Infection with human T-cellleukemia virus type 1 in patients with leukemia. N Engl J Med 1988; 318: 219-22.

4. Okochi K, Sato H, Hinuma Y. A retrospective study on transmission of adult T cellleukemia virus by blood transfusion: seroconversion in recipients. Vox Sang 1984;46: 245-53.

5. Evatt BL, Stein SF, Francis DP, et al. Antibodies to human T cell leukaemiavirus-associated membrane antigens in haemophiliacs: evidence for infectionbefore 1980. Lancet 1983; ii: 698-701.

6. Saxinger C, Gallo RC. Application of the indirect ELISA microtest to the detectionand surveillance of human T-cell leukemia lymphoma virus (HTLV). Methods LabInvest 1983; 49: 371.

7. Agius G, Biggar RT, Alexander SS, et al. Human T-lymphotropic virus type 1antibody patterns: evidence of difference by age and risk group. J Infect Dis 1988;158: 1235-44.

HTLV-I IN ENGLISH PATIENTS

SIR,-In the past 30 months sera referred to us from 65 patientshave been found to have antibodies to human T-cell lymphotropicvirus type I (HTLV-I). Anti-HTLV-I was detected by gelatinparticle agglutination (Serodia ’ATLA’), ELISA (Abbott, DuPont), competitive and IgG capture radioimmunoassay, andindirect immunofluorescence. All weakly and some highly reactivesera were tested by western blot (Biotech). Those giving positivereactions in all assays were reported as anti-HTLV-I positive. Thepossibility of false-positive results caused by cross-reacting HTLV-II antibodies was not excluded.

All the patients were seen at English hospitals. Tropical spasticparaparesis (TSP) was diagnosed in 39 (60%) and adult T-cellleukaemia/lymphoma in 14 (22%). 29 (74%) of the TSP patientswere female (table). Of the 60 patients whose ethnic origin wasgiven, 55 (92%) were West Indian, 2 were of English descent, andthere was 1 Japanese, 1 Brazilian, and 1 Iranian.

HTLV-I RELATED DISEASE IN ENGLISH PATIENTS

*1 husband, I wife, I daughter. tIncludes 2 blood donors.

TSP is the clinical effect of HTLV-1 most frequently identifiedin the UK, and most UK patients infected with HTLV-1 seem to beof West Indian origin. However, the pattern of carriage of HTLV-1may not be the same as the pattern of disease, and unbiasedinformation is needed on the prevalence of HTLV-I (and ofHTLV-II) infection in the community. Interventions to preventspread of these viruses by transfusion, intravenous drug use, sexualcontact, and mother-child transmission will depend upon thesedata.

PHLS Virus Reference Laboratory,London NW9 5HT

JENNIFER H. C. TOSSWILLJOHN V. PARRY

GENE AMPLIFICATION TO DETECT FETALNUCLEATED CELLS IN PREGNANT WOMEN

SIR,-The placenta is thought to be an effective barrier againstthe transfer of cells, but several attempts have been made to detectfetal nucleated cells in the maternal circulation. In some studies

putative fetal lymphocytes were identified by the detection of theY chromosome in nuclei1,2 but there is a risk of artifacts since

presumptive 46,XY cells have also been observed in non-pregnantfemales3,4 and in blood from mothers later delivered of a femaleinfants5 Claims that fetal lymphocytes expressing paternally-derived HLA antigens can be readily detected in maternal venousblood have not been confirmed.3 Flow cytometry has revealed asmall number of cellular elements expressing an antigen (H315),produced by the syncytiotrophoblast, in the blood of pregnantwomen.6,7 However, most of these cells are of maternal origin.’We have used the polymerase chain reaction (PCR) to investigate

the incidence of nucleated cells containing the Y chromosome invenous blood samples from pregnant women.Blood was collected in heparin from males and non-pregnant

females and from pregnant women. Nucleated cells were separatedand stained by a monoclonal antibody to H315;6 the H315-positivecellular element was isolated by ’Dynabeads M-450’ (Dynal,Norway). For PCR we used Taq polymerase (Cetus) and

oligonucleotides Y1.1 and Y1.2 encompassing 149 base pairs of arepeated sequence of the Y chromosome.8 All investigations hadethical committee approval and the verbal permission of thoseinvolved.

Analysis by PCR gene amplification ofY-specific sequence.Lanes 2-4 = chorionic villi from male fetuses; lane 5 = lymphocytes from

male; lanes 6-8 = single 46,XY lymphocytes mixed, respectively, with 35,70,and 100 x 103 lymphocytes from non-pregnant female; lane 9 = DNA from100 x 103 lymphocytes from non-pregnant female; and lane 10 = DNA from asingle 46AY lymphocyte.

Amplification performed with primer Y 1.1 and Y 1.2: asterisk indicates thefragment corresponding to the Y-specific sequence.