Immunochemical studies in Wilson's disease.

The present study deals with a total of 28 cases of Wilson's disease, 50 normal individuals alongwith siblings and parents of eight cases. Male predominance (18 out of 28 cases), a median age of 11 years and universal presence of Kayser-Fleischer (K. F.) ring marked the cardinal features. Furthermore, 11 patients had hepatic-neural presentation while two had only the K. F. Ring without clinical abnormality. Single Radial Immunodiffusion (SRID) as the absolute quantitative procedure revealed a profound deficiency of ceruloplasmin with the levels ranging anywhere between 0.5 mg/dI to 23 mg/dI amongst the patients of Wilson's disease. The data from siblings and parents also revealed deficiency in 15 out of 23 serum samples when subjected to disc electrophoresis-benzidine screening procedure.

Rapid detection of enteric fever by coagglutination and countercurrent immunoelectrophoresis.

A total of 463 patients clinically suspected of enteric fever and 100 healthy individuals were investigated by coagglutination (COAG) and countercurrent immunoelectrophoresis (CIEP) for rapid diagnosis of enteric fever. The S. typhi was grown in blood culture in 32 (6.91%) and Widal test was positive in 126 (27.21%) cases. The serum COAG with local antiserum was positive in 165 (35.64%), serum COAG with standard antiserum in 163 (35.21%), Blood culture supernatant (BCS) COAG in 153 (33.05%), serum CIEP for antigen (Ag) in 118 (25.09%), BCS CIEP in 99 (21.39%) while serum CIEP for antibody (Ab) was positive in 34 (7.34%) cases. Only two healthy controls revealed positive COAG result with local antiserum. The sensitivity of all antigen detection tests was 100% except BCS CIEP (98.25%) in the first week of fever and declined rapidly to 75.79% for serum COAG tests, 69.47% for BCS COAG and dramatically to 37.89% for serum CIEP for Ag and 22.11% for BCS CIEP tests during the second week whereas the sensitivity of serum CIEP for Ab detection rose from 17.54% to 23.16% from first to second week of illness. In view of the resulting data, it is suggested that both COAG and CIEP may be employed for the rapid diagnosis of enteric fever in the routine clinical setup.

Comparative evaluation of immunochemical methods for estimation of albumin in microalbuminuria.

Three immuno assays namely radioimmunoassay (RIA), radial immunodiffusion (RID) and rocket immunoelectrophoresis (RIE) were compared for their performance and utility. The accuracy limits of the methods were compared and also between methods using RIA as the reference. Urine samples, from known diabetic patients with albumin concentration ranging from 2.5 mg/l to 120 mg/l were analysed by the three methods. The mean differences were only 0.91 mg/dl and 0.5 mg/dl respectively for RID vs RIA and rocket vs RIA which is not statistically significant. Excellent correlation was seen between RIA and RIE (r = 0.98) and also between RIA and RID (r = 0.97). Compared to RID, RIE required less time and was more precise. RIA is suited for assaying large sample loads yet not suited for laboratories receiving samples occasionally. For a small pathological laboratory with limited facility rocket electrophoresis may be the most suitable method taking into consideration accuracy, time and cost.

Immunochemical studies in multiple myeloma.

The study deals with a total of 72 patients with Plasma cell dyscrasias (PCD) selected on the basis of atypical plasmacytosis in the bone marrow aspirate and radiological evidence of osteolytic lesions. Males(48) outnumbered the females (24). Pathological fracture and paraplegia was the commonest presenting symptom encountered in 38 patients. Electrophoresis of serum for 'M' band and Immunoelectrophoretic analysis of the serum revealed IgG myeloma in 40 patients followed by, IgA myeloma(13), Light chain disease (12) and other variants in remaining seven cases. The urinary Bence Jones proteins were detected in a total of 34 cases and was frequently encountered with IgA myeloma (7 out of 13) compared with IgG myeloma (13 out of 40) when analysed in Disc electrophoresis. Kappa light chain was observed in 21 cases and lambda counterpart in nine cases without any clinical significance. One case of solitary myeloma terminated in characteristic multicentric multiple myelomatosis within a span of six months in the sequential follow up study. We recommend the triangular approach to diagnosis of paraproteinemia with a special emphasis on immunoelectrophoresis for typing multiple myeloma and allied disorders along with disc electrophoresis for the demonstration of urinary Bence Jones protein in the routine set up.

Bisalbuminemia: an interesting electrophoretic abnormality--a report of two cases.

We hereby report two cases of Bisalbuminemia for its extreme rarity in Indian literature. In an attempt to look for 'M' component, polyacrylamide gel disc electrophoresis of serum displayed observation. The saline eluates of the slower (variant in our cases) and the faster component showed a reaction of complete identify in the agar gel double diffusion test when tested against monospecific antialbumin antibody. The immunoelectrophoretic analysis also confirmed the antigenic similarity between the two fractions. A wider use of disc electrophoresis perhaps would disclose additional cases of Bisalbuminemia in normals and in disease states.

Analysis of immunoglobulin deficiency cases: a five year study.

A total of 734 serum specimens from various clinical disorders along with 100 control samples from healthy subjects were processed for estimation of serum IgG, IgA and IgM employing single radial immunodiffusion procedure. Immunoglobulin deficiency, either selective or combined was noted in 31 males and 24 females in all age groups. Of the 55 cases encountered it was secondary immunoglobulin deficiency which was seen on a larger scale and encountered in patients with Multiple myeloma (16 out of 32) followed by Leprosy (14 out of 250), Lymphoma (5 out of 43), Malaria (4 out of 137), Burns (4 out of 52), Rheumatoid arthritis (2 out of 69) and non lymphoreticular malignancies (1 out of 41) in decreasing order of frequency. Primary immunoglobulin deficiency was observed in nine cases comprising of six belonging to Idiopathic late onset immunoglobulin deficiency, two of dysgammaglobulineamia and a solitary case of Ataxia telangiectasia. Panimmunoglobulin deficiency was observed in six cases, 11 had a dual deficiency while 38 showed deficiency of an isolated class with selective IgA deficiency in 20 cases. Furthermore, one patient each had total absence of IgG or IgA while IgM was not detectable in seven patients. A high suspicion index along with a regular rapport between the clinician and the laboratory personnel is necessary in the diagnostic set up of immunoglobulin deficiency states.

Extended spectrum beta-lactamase mediated resistance to third generation cephalosporins in Klebsiella pneumoniae in Nagpur, central India.

Out of 66 clinical isolates of Klebsiella pneumoniae, 17 showed resistance or decreased susceptibility to third generation cephalosporins (17 to cefotaxime, 16 to ceftriaxone, and 9 to ceftazidime) while the remaining 49 were sensitive by the disc diffusion method. The minimum inhibitory concentrations (MICs) of the third generation cephalosporins (3GC) for the strains ranged from 2-128 micrograms/ml by agar dilution method. Their sensitive phenotypes had zone diameters smaller (mean difference 3. 1 mm for ceftriaxone, and 6.5 mm for ceftazidime), and MICs > 10 fold higher than the corresponding values in the fully sensitive isolates. Resistance to cefotaxime was transferred to recipient Escherichia coli K12 strain in 15 isolates. All the resistant isolates were sensitive to imipenem but were variably sensitive to aminoglycosides, and quinolones. In all 17 resistant isolates extended spectrum beta-lactamase (ES beta L) was detected. The sensitivity testing systems may fail to recognise the potential ES beta L mediated resistance to 3GC. Hence ES beta L detection should be routinely undertaken.

Haemolytic Escherichia coli in etiopathogenesis of pyelonephritis in rats--an experimental study.

Of the 26 urinary E. coli strains employed in the present study, 16 were haemolytic and 10 were non haemolytic in character. Of the 16 haemolytic strains injected through the exposed femoral vein of the rats, 12 showed the lesions of pyelitis, pyelonephritis and proliferation of glomerular tuft while 2 showed only the lesions of pyelitis and pyelonephritis. It was interesting to note that none of the nonhaemolytic strains produced any lesion in the animal. The data demonstrated a positive correlationship between haemolytic activity of E. coli and pyelonephritis in rats.

Study of suspected plague cases for isolation and identification of Yersinia pestis.

A total of 62 suspected patients of plague were investigated for evidence of Yersinia pestis, by blood culture, lymph node aspirate culture, sputum culture, animal inoculation and serology for f1 antibodies against f1 antigen of Yersinia pestis. None of the samples was positive by direct smear examination and culture for Yersinia pestis, as well as for serology. The non positivity of the cultures is discussed.

Species distribution of clinical isolates of Staphylococci.

Species identification of 103 strains of staphylococci isolated from various clinical specimens revealed as many as eight species. S. aureus was the commonest isolate (57.28%) followed by S. epidermidis (17.47%). S. hominis (8.73%), S. saprophyticus (6.79%), S. intermedius, S. capitis and S. cohni (1.94% each) in decreasing order. They were mostly isolated from pus (41.74%) followed by urine (22.33%) and blood (18.44%) accordingly. Of the total isolates, 65 were coagulase positive while 38 isolates were coagulase negative. Phase typing of S. aureus strains revealed that 49 isolates were typable of which 17 isolates belonged to phase group III, 16 to phase group 1, 4 to group II and 3 belonged to group V while 9 strains could not be grouped. The antibiotic resistance pattern showed maximum resistance to penicillin (63.10%) while resistance was minimum with gentamycin (8.73%). Resistance to ampicillin (51.45%), cephazoline (51.45%), erythromycin (31.06+) and cloxacillin (18.44%) was low to moderate. The increasing recognition of pathogenic potential of various species of staphylococci and emergence of drug resistance amongst them denotes the need to adopt better laboratory procedures to identify and understand the diversity of staphylococci isolated from clinical material.