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At least 19 recordsLinked to original sources
PubMed1978

Isolation of tuberculin peptides from tuberculin purified protein derivative (PPD).

Tuberculin purified protein derivative (PPD) obtained from the filtrate of Mycobacterium tuberculosis was hydrolysed with proteinase, trypsin, or chymotrypsin. Each hydrolysate consisted of a tuberculin peptides mixture (TPM). From each TPM 16 fractions were obtained by ion-exchange chromatography on Dowex 50W-X8 but only one fraction was isolated from each of the 16 fractions which showed tuberculin activity in guinea pigs sensitized with M. bovis (bcg) or M. tuberculosis. This fraction was designated "purified tuberculin peptide" (PTP). The PTP fraction from the proteinase hydrolysate (PTP-proteinase) was rechromatographed on Dowex 1-X2 and two tuberculin peptide fractions having molecular weights of 3200 and 12,000 were isolated. The potency of these two fractions was assessed in guinea pigs sensitized with M. bovis (BCG) and with M. tuberculosis and they were approximately 4 to 7 times more potent than either the international standaCG and of at least equal potency to either PPD-S or Connaught PPD in guinea pigs sensitized with either M. kansasii, M. scrofulaceum, M. intracellulare, or M. avium whereas very little if any cross-reactivity was elicited by these two fractions. This lack of response indicates that either fraction could be used as an aid to differentiate between sensitization due to M. tuberculosis or M. bovis and sensitization attributed to other mycobacteria.

Animals↗
PubMed1978

Comparison of the tine and Mantoux tuberculin tests. Report of the Tuberculin Subcommittee of the Researched Committee of the British Thoracic Association.

Intracutaneous (Mantoux 5 TU) and tine tuberculin tests were performed on the opposite arms of 307 people. The results of each test were read by two independent observers at either 48 or 72 hours. Positive readings were recorded in 59% of the Mantoux tests; induration was 10 mm or more in 34.7% of cases. Positive readings were recorded for 3.9% of the tine tests, and a further 15.5% were recorded in the doubtful category. The tine test is unsuitable for epidemiological use because of the high proportion of negative and doubtful results in people positive on the Mantoux test. For the same reasons its usefulness in clinical practice is very limited.

False Negative Reactions↗
PubMed1975

The multi-facets of tuberculin standardization.

Some parameters affecting the standardization of a tuberculin preparation have been evaluated and it has been shown that: 1) The loss of tuberculoprotein on the surfaces of containers is in the order of 0.15 mug/cm2 and the 0.18 mug/cm-2 for glass and plastic surfaces respectively, and that this loss can be prevented by the addition of a nonionic anti-adsorption agent such as Tween 80. 2) For non-stabilized tuberculin PPD solutions the losses of PPD due to adsorption increased linearly with the Surface to Volume ration. 3) Exposure to light had a deleterious effect on tuberculin PPD decreasing its biological potency and altering its appearance. This effect increased with the dilution of the tuberculoprotein solution, and can be prevented by storing the tuberculin in amber containers. 4) The potency of tuberculoprotein was dependent on its molecular weight, and the % distribution of tuberculoprotein molecules of different molecular weight varied from batch to batch. This variation can be eliminated by preparing a large batch of tuberculin in a dried form sufficient to fill the demand for many years to come. 5) The route of injection and the species or strains of mycobacteria used for sensitization of guinea pigs affected the relative potency of a tuberculin preparation. 6) The size of a tuberculin skin reaction increased as the volume of tuberculin injected intracutaneously into sensitized guinea pigs was increased although the number of tuberculin units administered was the same. 7) The sizes of skin reactions to certain doses of tuberculin was affected by other doses of tuberculin being injected simultaneously at other sites. 8) Sex and age of guinea pigs affected the size of the tuberculin skin reaction. 9) The genetic make-up of the test animals affected the relative potency of a given tuberculin preparation. 10) Racial differences among various populations might account for different levels of tuberculin reactivity to a given tuberculin preparation. It is hoped that this work has made us aware of the many problems which can be encountered in standardizing tuberculin and it is also to be hoped that the solutions we have offered to some of these problems can be useful to those attempting to standardize allergens other than tuberculin.

Animals↗
PubMed1979

Tuberculin test conversion during repeated skin testing, associated with sensitivity to nontuberculous mycobacteria.

To determine whether repeating the tuberculin test after a brief interval might result in tuberculin conversion, we tested 213 healthy volunteers twice, 1 month apart, with 5 TU of tuberculin purified protein derivative (PPD). Three nontuberculous mycobacterial antigens (PPD-G, PPD-Y, and PPD-B) were also applied with the first tuberculin test. By widely used criteria, 14 volunteers (6.6 per cent) converted their tuberculin tests from negative to positive on the second testing. Whereas 13 of 103 subjects (12.6 per cent) with nontuberculous antigen sensitivity converted their tuberculin test to positive, only one of 110 subjects (0.9 per cent) with no known prior mycobacterial sensitivity converted to positive (P less than 0.005; x2 = 10.05). When retested with 5 TU of tuberculin PPD 6.5 months after the second test, nine of 13 converters reverted to negative. We conclude that tuberculin conversion may occur when the skin test is repeated at 1 month, and that boosting of cross-reacting mycobacterial sensitivity might have caused a portion of the conversions in this population of young, healthy midwestern volunteers. Sensitivity to Mycobacterium tuberculosis might also be boosted by tuberculin testing. Because the prevalence of sensitization by tuberculous and nontuberculous mycobacteria can be expected to vary in different populations, the significance of tuberculin conversion will also vary with the population being tested.

Adolescent↗
PubMed1975

Comparison of the specificty of human and bovine tuberculin PPD for testing cattle. 1--Republic of Ireland.

A tuberculin testing trial in cattle was carried out in the Republic of Ireland to compare the specificity for bovine tuberculosis of a human purified protein derivative (PPD) tuberculin (Weybridge) with that of a bovine PPD (Rotterdam), and to determine whether discrimination between specific and non-specific reactions to mammalian tuberculin is better with doses of tuberculins smaller than those traditonally used for testing cattle. Tests were carried out in 510 cattle, 395 of which were shown by post mortem examination to be tuberculous and 115 non-tuberculous. Three dilutions at five-fold intervals of both mammalian tuberculins were used together with two dose levels of avian tuberculin PPD (Weybridge), and all reactions were measured both by increase in skin fold thickness and by diameter of induration. In the environment of this trial, the bovine PPD was shown to be more specific for bovine tuberculosis than the human PPD, and particularly in differentiating from "skin tuberculosis". There was no indication of greater specificity at lower doses of tuberculin. Measurement of induration diameter proved a satisfactory alternative method of reading tuberculin reactions in cattle under field conditions.

Animals↗
PubMed1977

Multiple skin testing of tuberculosis patients with a range of new tuberculins, and a comparison with leprosy and Mycobacterium ulcerans infection.

Four hundred and seventy tuberculosis patients were each skin tested with four of a range of 17 mycobacterial reagents in four countries in all of which tuberculosis and leprosy were endemic. Sixteen of the reagents were new tuberculins prepared from extracts of living mycobacteria disrupted by ultrasonic disintegration and the last was PPD, RT23.The effect that tuberculosis exerted on the delayed-type skin test response to these antigens was assessed by comparing results for tuberculosis patients with those for Tuberculin positive and Tuberculin negative control populations. Tuberculosis patients on Rifampicin therapy showed no difference in their skin test responses to any of the antigens from those patients on other forms of antituberculosis treatment.Amongst the normal population it was found that possession of Tuberculin positivity was associated with an enhanced response to all the other mycobacterial antigens with the exception of A(*)-in which demonstrated a reciprocal relationship with Tuberculin in Burma. It was also noted, in Burma particularly, that sensitization to mycobacterial species other than Mycobacterium tuberculosis, especially to the slow growers, plays a role in determining responses to different mycobacterial species.In tuberculosis patients enhanced skin test responses were also seen but only in those countries, e.g. Libya, where the prevalence of mycobacterial species was low. Where mycobacteria were common, as in Burma, the converse was true and tuberculosis was associated with a diminished skin test response to each antigen. The high prevalence of A(*)-in positivity in Burma, its reciprocal relationship with Tuberculin there and the results for all the antigens in the tuberculosis patients indicate that the cell mediated skin test response may have a threshold. If this is exceeded the skin test becomes negative so that non-reactors then include those who have been excessively sensitized as well as those who have not been sensitized. Despite this, a greater percentage of tuberculosis patients in each country responded to the specific reagent Tuberculin than did the control populations and their mean positive induration sizes were consistently larger. Nevertheless, amongst the tuberculosis patients in Burma 13% were complete non-reactors to Tuberculin and this apparent anergy also applied to the other reagents with which these individuals were tested.This differs from lepromatous leprosy where the anergic state pertains exclusively to M. leprae and a few seemingly closely related species. The breadth of anergy in M. ulcerans infection has not been measured but it is known to effect both Burulin and the PPD, RT23.Just as in leprosy and M. ulcerans infection, tuberculosis can be shown to have a disease spectrum here detected by multiple skin testing. The significance of this spectrum and its similarities with and differences from that of the other mycobacterioses is discussed.

Adolescent↗
PubMed1975

Comparison of the specificity of human and bovine tuberculin PPD for testing cattle. 2. South-eastern England.

A tuberculin testing trial was carried out in eight counties of south-eastern England to compare the specificity for bovine tuberculosis of Weybridge human PPD with that of Rotterdam bovine PPD. The matching of these two tuberculins for potency in naturally infected cattle had already been established, the bovine PPD being approximately one-and-a-half times more potent than the human PPD per unit of weight. In 1110 cattle in 25 herds with histories of long-standing freedom from tuberculosis and in which non-specific tuberculin sensitivity was present, cross reactions were less to the bovine PPD than to the human PPD, showing that in the environment of this trial the bovine PPD was more specific than the human PPD. Induration diameter was a satisfactory alternative to skin thickening as a measure of tuberculin reactions in cattle under field conditions. Due to the steep slope of the dose-response curves of the avian PPD in the different groups of non-tuberculous cattle, the discriminating power of the comparative test, using avian and mammalian tuberculins, was less at lower doses of tuberculin. Concentrations of 1-0 mg per ml of bovine PPD and 0-5 mg per ml of avian PPD are recommended for use in a comparative tuberculin test.

Animals↗
PubMed1976

Potency of purified tuberculin determined by the shock method on hamsters.

Two lots of purified protein derivative of tuberculin with different potencies determined by skin tests on guinea pigs, were examined for their capacity to induce shock in hamsters sensitized with BCG. Groups of hamsters were injected with varying doses of a highly potent strain of BCG grown in Dubos fluid medium. 4-5 weeks later two groups of animals sensitized with corresponding doses were injected intraperitoneally with 1 mg of each lot of tuberculin. 12 of the 20 hamsters given the strong tuberculin died, while all those given the weak tuberculin survived. An attempt was made to evaluate the mutual potency of the two lots of tuberculin by injecting groups of sensitized animals with 1 mg strong or 4 mg weak tuberculin. The difference in the number of deaths from shock in the two groups was not significant.

Animals↗
PubMed1978

Evaluation of the tuberculin test in Malawi.

In a trial involving 371 head of cattle, the results of using avian plus bovine tuberculin and avian plus mammaliam tuberculin were compared. One group of 185 cattle from areas of low tuberculosis incidence and another group of 186 cattle from areas of high incidence were tuberculin tested and examined for pathological lesions. Using the interpretation "non-specific infection not established" with avian plus mammaliam tuberculin on the readings from cattle from high incidence areas, the test detected 83% of the cattle with visible lesions, whereas the avian plus bovine tuberculin test only detected 63%. The avian plus mammaliam tuberculin test is therefore more accurate; however, it appears to be too sensitive as it produces a higher rate of inconclusive results from non-infected cattle, and is thus more difficult to interpret. Many cattle giving positive reactions were found to be free of tubercular lesions when examined post mortem; the possibility is discussed of their allergy to the test being caused by mycobacteria other than tuberculosis, as mycobacteria belonging to Runyons Group IV have been isolated from some of these cases.

Animals↗
PubMed1977

Evidence for transmission of lymphocyte responses to tuberculin by breast-feeding.

The possibility that cell-mediated immunity could be acquired by breast-feeding was evaluated in a prospective study of 26 tuberculin positive and 9 negative puerperal mothers and their infants. 13 infants of the positive and all the infants of negative mothers were breast-fed. Tuberculin-reactive T cells were found in colostrum and early milk of most positive but in none of the negative nursing mothers. A significant number (8/13) of infants born to positive mothers had tuberculin-reactive peripheral blood T cells after 4 weeks of breast-feeding compared with bottle-fed infants (1/13) of positive mothers or breast-fed infants (0/9) of negative mothers. Examination of cord blood for tuberculin-reactive T cells provided no significant evidence of transplacental transmission of responsiveness to tuberculin. The results suggest that breast-fed infants may passively acquire T cell responsiveness to a specific antigen by ingestion of breast milk.

Blood↗
PubMed1975

Comparison of the specificity of human and bovine tuberculin PPF for testing cattle. 3. National trial in Great Britain.

A field trial on a country-wide basis was undertaken to compare the specificity for bovine tuberculosis of single and comparative tuberculin tests in cattle using either Weybridge human or Weybridge bovine PPD. The tests were made on 10,305 cattle in 179 herds distributed throughout all regions of England, Scotland and Wales. Results showed that a comparative tuberculin test using avian PPD with either human or bovine PPD had a much higher efficiency than a single injection of mammalian tuberculin in the neck of cattle, and confirmed that a comparative test is still essential in the British environment. Weybridge bovine PPD gave significantly better discrimination between tuberculous and non-tuberculous cattle than Weybridge human PPD when used together with avian PPD in a comparative tuberculin test. The diameter of induration gave an absolute measure of the extent of oedema, if present, and induration diameter used in conjunction with skin thickening increased the sensitivity and specificity of the test. Rules of interpretation were developed and are presented for an intradermal comparative tuberculin test in cattle using Weybridge avian and bovine PPDs.

Animals↗
PubMed1978

Stability of dilute solutions of tuberculin purified protein derivative.

The biological potency of 15 commercially available tuberculin solutions prepared from a master batch of tuberculin purified protein derivative (PPD) (PPD-CT68) and using a newly formulated diluent (Connaught diluent) containing 0.0005% Tween 80 as an anti-adsorption agent and 0.3% phenol as a preservative, was determined after storage for various intervals at 4, 24 and 37 degrees C. The 5 tuberculin units (TU) per 0.1 ml dose solutions were bioequivalent to a non-stabilized solution of PPD-S whereas the 1 TU and 250 TU per dose solutions were equivalent by calculation to a non-stabilized solution of PPD-S. It was found that the PPD solutions of all 3 strengths, 1, 5 and 250 TU per dose, were stable for at least 3 years at 4 degrees C and for 2 years at room temperature (24 degrees C). Even at 37 degrees C the solutions of all 3 strengths were stable for at least 1 year. The stability of Connaught tuberculin PPD solutions has not been affected by the changes in formulation. The stability data suggest that the expiry date of the newly formulated tuberculin products could be at least two years from the data of the last satisfactory potency test. Although these products are stable for at least 1 year even at 37 degrees C, we nonetheless agree with the Canadian and U.S. regulations that they be stored at 2 to 8 degrees C in their original containers.

Animals↗
PubMed1978

Pathogenicity for cattle of atypical mycobacteria isolated from feral pigs and cattle and the correlation of lesions with tuberculin sensitivity.

Two experiments involving the inoculation of cattle with atypical mycobacteria are described. In the first experiment groups of 5 cattle were inoculated either subcutaneously or into a mesenteric lymph node with a strain of M. scrofulaceum or M. intracellulare. Four weeks and 10 weeks after inoculation the cattle were tuberculin tested with bovine PPD, avian PPD and homologous PPDs. The pathological changes observed were similar within each group of cattle inoculated with the same strain of mycobacteria. A significant interaction was demonstrated between the strain and the route of inoculation. In the second experiment 17 cattle were similarly inoculated by either of the two routes with 1 of 6 strains of M. intracellulare, a strain of M. scrofulaceum or a strain of Runyon Group IV, all of which had been isolated from feral pigs, or a strain of M. intracellulare of bovine origin. Tuberculin tests were carried out after 4 weeks and 10 weeks. Only the isolate from a bovine lymph node produced a significant level of sensitivity to bovine PPD. Cultural isolation of the mycobacteria from autopsy material was not correlated with the presence of macroscopic lesions nor with sensitivity to bovine PPD. The response to bovine PPD of cattle infected with these atypical mycobacteria decreased between 48 h and 96 h after injection of the tuberculins. As the maximum difference in the response to bovine and avian tuberculins occurs at 72 h a comparative tuberculin test should be read at this time to eliminate non-specific reactors.

Animals↗