A pilot study of HTLV-I infection in high-risk individuals & their family members from India.

BACKGROUND & OBJECTIVES: Human T lymphotropic virus-I (HTLV-I) has been associated with adult T cell lymphoma/leukemia (ATLL). There are Indian studies on HTLV-I infection among people with sexually transmitted infection, but no large study has been conducted on individuals with haematological malignancies. In this group of individuals, serology is known to under-diagnose HTLV-I infection. This study was carried out to identify serologically and where possible with molecular techniques, HTLV-I infection in individuals with haematological malignancies. To understand the modes of transmission, family members of individuals with proven HTLV-I infection were also studied. Individuals with sexually transmitted infection (STI), blood donors and pregnant women were also studied. METHODS: Particle agglutination test was used to detect antibody to HTLV-I. HTLV genome was amplified by polymerase chain reaction (PCR) and detected with probes by digoxiginin (Dig) ELISA. RESULTS: There was no serological evidence of HTLV-I infection among the healthy blood donors and pregnant women studied. High prevalence of anti-HTLV-I antibody was identified in the patients with haematological malignancies (8 of 86 patients, 9.3%) and a lower prevalence in individuals with STI (8 of 670 individuals, 1.2%). In the STI group, all 8 individuals seroreactive to HTLV-I were coinfected with human immunodeficiency virus (HIV). In the group with haematological malignancies, three of 22 (13.6%) patients with leukemia, 3 of 11 (27.3%) with cutaneous T-cell lymphoma (CTCL) and 2 of 53 (3.8%) with lymphoma were reactive for anti-HTLV-I antibody. In this group, PCR identified all the seroreactive individuals tested. There were also seronegative infected individuals who were only identified by PCR. There was also a large number of seronegative family members who were only positive by PCR. INTERPRETATION & CONCLUSION: The study revealed a strong disease association of HTLV-I with haematological malignancies and evidence for both horizontal and vertical transmission of the infection in the Indian population. HTLV-I infection appears to be common among family members of individuals with HTLV-I associated haematological malignancies.

NESTROFT as a screening test for the detection of thalassaemia & common haemoglobinopathies--an evaluation against a high performance liquid chromatographic method.

The NESTROFT method (Naked Eye Single Tube Red Cell Osmotic Fragility Test) was evaluated against a high performance liquid chromatographic (HPLC) method for its usefulness in screening for beta-thalassaemia and some of the common haemoglobinopathies. Blood samples (137) from patients with suspected haemoglobin disorders were analyzed by both methods. Among the 63 patients with heterozygous beta-thalassaemia on HPLC, NESTROFT was positive for 49, 'doubtful' for 13 and negative for 1. Of the 32 patients with other haemoglobin disorders, 28 were positive on NESTROFT and 4 were 'doubtful'. Of the 42 'normal' samples, NESTROFT was positive for 6, 'doubtful' for 8 and negative for 28. This test showed an overall sensitivity of 98.7 per cent, specificity of 66.6 per cent, positive predictive value of 87 per cent and negative predictive value of 96.5 per cent. We conclude that, NESTROFT is a suitable test for screening for beta-thalassaemia and the common haemoglobinopathies seen in India. It is easy to perform, simple, inexpensive and does not require sophisticated equipment. Subjects who are NESTROFT 'positive' or 'doubtful' deserve further investigation.

Prenatal diagnosis of beta-thalassaemia mutations using the reverse dot blot technique.

BACKGROUND: Beta-thalassaemia is the most common genetic disorder among Indians and a number of mutations causing this disease have been reported. Since effective treatment of thalassaemia major is complicated and very expensive, prenatal diagnosis has become an important option for those at risk of having an affected foetus. We report the use of a rapid hybridization method called 'reverse dot blot' for detection of specific mutations of the beta-globin gene. METHODS: DNA was obtained from a 12-week-old foetus by chorionic villus sampling and was amplified using specific primers by the polymerase chain reaction and analysed by the reverse dot blot test. Results were available within 36 hours after sampling. RESULT: The father and mother were found to be heterozygous for codon 15 (G-A) mutation of the beta-globin gene. The foetus was normal. CONCLUSION: Reverse dot blot is a rapid and reliable technique for mutation detection in the beta-globin gene and can be useful for antenatal diagnosis.

Comparison of HPLC & EMIT methods of cyclosporine assay in blood after bone marrow transplantation.

Cyclosporine (CsA) analysis in blood from patients who had undergone bone marrow transplantation for various haematological disorders was done both by high performance liquid chromatography (HPLC) and enzyme multiplied immunoassay technique (EMIT) and the results were compared. HPLC kit from Biorad Laboratories, USA, and EMIT kit from SYVA, UK, were used. The procedure for EMIT was slightly modified in-house to suit the Hitachi 704 discrete selective analyser. The CsA values obtained by these two methods correlated well within the therapeutic range (r value 0.96), HPLC method being most suitable outside the therapeutic range. Although HPLC is the ideal method for CsA, EMIT is quite suitable and can be adopted by any laboratory with an autoanalyser incorporating our modified procedure.

Treatment of childhood acute lymphoblastic leukaemia with the BFM regimen.

Thirty-eight children with acute lymphoblastic leukaemia were treated with the BFM regimen. Thirty-six (94.7%) achieved complete remission (CR). Twenty 58.8%) of 34 evaluable patients are in continuous complete remission (CCR) at a median follow-up of 33 (range 19-81) months. Long-term disease-free survival was better in the 2-9 years age group (83%) when compared to the 10-14 years group (43%) (P < .05).

Treatment of acute lymphoblastic leukaemia in adults with modified BFM regimen.

Sixty-two adults with acute lymphoblastic leukaemia (ALL) were treated with a modified BFM regimen. Forty-two (70%) achieved complete remission (CR). Twenty-one percent of all evaluable patients and 32.4 percent of complete responders are in continuous complete remission (CCR) at a median follow-up of 41 months (range 24-81 months). Long-term survival was better in T- ALL (47.1%) when compared to precursor-B ALL (4.8%) (P < 0.04).

Transfusion associated HIV infection in patients with haematologic disorders in southern India.

Seventy patients with congenital coagulation disorders (group A) and 202 other patients (group B) attending the Haematology clinic at the Christian Medical College and Hospital, Vellore (India) were screened for HIV infection between March 1989 and April 1991. Fifty five patients in group A and 131 patients in group B had received blood or blood products in the past. Nineteen transfused patients (9 in group A and 10 in group B) had received blood or blood products exclusively from the hospital blood bank and none of them was HIV infected. Among the remaining 167 transfused patients, 14 (30.4%) of the 46 patients in group A and 6 (4.9%) of the 121 patients in group B were found to be positive for HIV. In group A, 13 of the 14 infected patients had received commercially available cryoprecipitate which was thus found to be the most frequent source of infection. In group B the source of infection was most probably unscreened HIV infected blood which was transfused.

Surgery in patients with congenital coagulation disorders.

BACKGROUND. Surgery is occasionally necessary in patients with congenital coagulation disorders. Major surgery for patients with haemophilia was not being done in India until recently. This paper reports the experience of a single referral centre. METHODS. The data of 52 patients who were operated upon were collected from the hospital records retrospectively between 1984 and 1986 and prospectively thereafter. They included the surgical procedure performed, replacement therapy used and complications encountered. RESULTS. Fifty-nine procedures were performed of which 26 were major, 30 minor and 3 were diagnostic angiograms. Blood components produced in the hospital blood bank were commonly used for replacement and primary haemostasis was achieved in all patients. Delayed bleeding due to inadequate factor levels occurred in 12 procedures and was controlled by increasing the factor replacement. One patient died of suspected acute myocardial ischaemia. CONCLUSION. In India surgical procedures can be safely performed in patients with congenital coagulation disorders.

Developing hemophilia services in India.

With a population of 853 million there should be 51,204 patients with hemophilia A in India assuming a prevalence of 6/100,000 population. With the current birth rate of 32/1000, 1,300 new patients with hemophilia A will be born each year. Hospital based data suggests that hemophiliacs in India suffer from preventable morbidity because doctors do not know enough about the disease and its management, because laboratory diagnostic facilities are inadequate and because there is not enough therapeutic material or even if it is available the patients do not have the resources to purchase it. This article reviews the current status of hemophilia in India and suggests measures to improve hemophilia services within the health care infrastructure available in the country.