Immunofluorescence testing for chlamydial antibodies.
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Publications and source records attributed to S Darougar.
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The relationship of three infectious agents, Chlamydia, cytomegalovirus, and Yersinia enterocolitica, to the aetiology, clinical course, and diagnosis of Crohn's disease was studied. There was no evidence of chlamydial infection in Crohn's disease and no indication either that cytomegalovirus alters the outcome of acute attacks or that infection with Y. enterocolitica is being misdiagnosed as Crohn's disease.
We present the case of a 21-year-old girl with a uniocular juxtapapillary choroiditis. During the course of her illness the titre of antichlamydial IgG increased from 1/32 to 1/4096 against Chlamydia trachomatis TRIC serotypes J and C, and antichlamydial IgM appeared in her blood. Toxoplasma dye test was positive at a level of 1/128 but no increase in the titre of antibody was detected during the course of her infection. The relevance of these findings to her ocular lesion is discussed.
The simplified one-passage technique of culture in irradiated McCoy cells, in conjunction with certain other developments in technique, was used to isolate Chlamydia trachomatis from specimens collected from 78 children with trachoma in Douz, Southern Tunisia. The results show that C. trachomatis is not confined to the upper tarsal area of the conjunctiva in hyperendemic trachoma. The higher isolation rate and corresponding increase in the number of inclusions obtained from swabbings of the upper fornix and lower lid in addition to the conventional collection from the upper tarsus show the superiority of collecting specimens for culture from the whole conjunctiva. Specimens could be collected from the whole conjunctiva by using 1 swab for each eye and pooled for subsequent inoculation, so that the laboratory incurred no additional work. A close correlation was observed between isolation rate, together with the number of inclusions, obtained in cell culture, and intensity of inflammatory disease in hyperendemic trachoma. The sensitivity and practicability of this cultural test should provide a valuable laboratory index for use in epidemiological and therapeutic studies of trachoma.
Shedding of Chlamydia trachomatis in the eye secretion (tears) of patients with either hyperendemic trachoma or paratrachoma was studied. The method of collection of eye secretion with cellulose sponges is proved to be simple, faster, and more practicable and yielded a higher rate of chlamydial isolation than aspiration. The chlamydial isolation rates in eye secretion in chlamydia-positive paratrachoma patients in London or trachoma patients in Iran was 84 and 49% respectively. It was found that the chlamydial isolation rate from eye secretion is directly related to the number of inclusions present in the conjunctival swabbings. The results of this study indicated that patients with moderate to severe hyperendemic trachoma or paratrachoma are the main reservoir of infection. In the developing countries of the Middle East and Africa the shedding of chlamydia in the eye secretion of persons with these diseases is a major factor in the transmission of them by means of flies, fingers, towels, or bed clothes.
The sensitivity of human embryonic kidney (HEK) cell culture and the complement fixation test (cft) in the diagnosis of adenovirus ocular infection has been compared. The optimum time for collection of specimens to obtain the best results by each test has been examined. Sixty-one (53%) of 116 patients with moderate to severe follicular conjunctivitis, clinically indicative of adenovirus infection, had serological evidence of infection or had the virus isolated from ocular swabs. Virus was isolated from 53 patients (45%) and significant antibody rises were found in 33 (27%). When each test was carried out under optimum conditions, virus was isolated from 82% of cases and serological evidence of infection found in 72%. Serological responses as detected by the CFT appeared to be inferior in infections caused by adenovirus type 7 to those encountered in infections due to adenovirus types 3 and 8.
Recent isolation studies have shown Chlamydia trachomatis to be an important aetiological agent in acute salpingitis in women. The present serological study indicates that C. trachomatis is the probable aetiological agent in two-thirds of 143 women with pelvic inflammatory disease (PID). In general, high levels of chlamydial antibody were found in sera and fluids aspirated from the pouch of Douglas and such antibody titres were shown to correlate with the severity of clinically graded tubal inflammation.
In a double-blind comparison of two regimens of triple tetracycline (Deteclo, Lederle) in the treatment of nongonococcal urethritis, 68 (88.6%) of 70 patients treated with one tablet twice for 21 days and seen four weeks after starting therapy had satisfactory results. This was significantly better than the findings among the 73 patients treated with one tablet twice daily for seven days and followed for four weeks, among whom only 47 (64.4%) had satisfactory results. Results were also better for the group treated with the 21-day regimen at three months afer the start of treatment. When analysed individually at four and 12 weeks, urethral discharge, urethral Gram-stained smears, and first-glass urine test all gave similar results, which were markedly better than those before treatment. It appears that the longer course of treatment it indicated where any regular partner may not be treated. Slightly fewer patients had satisfactory results among those who admitted consuming alcohol than among those who did not. Chlamydiae-negative patients, treated for seven days, had fewer clinically satisfactory results than other sub-groups.
The role of Chlamydia trachomatis in pelvic inflammatory disease (PID) diagnosed without laparoscopy was assessed by measuring antichlamydial antibodies in the patient's serum and by comparing the results with those in patients with uncomplicated non-specific genital infection (NSGI) and gonorrhoea and in non-infected controls. A modified microimmunofluorescence test was used. Patients with severe PID had significantly more positive antichlamydial IgG and IgM results than did control subjects, patients with gonorrhoea, and patients with NSGI. Less severe PID was associated with significantly raised levels of antichlamydial IgG antibodies compared with NSGI and controls and with raised levels of IgM antibodies compared with controls. Two patients with PID had lower genital tract gonorrhoea, one of whom had raised antichlamydial antibody levels. These findings may indicate a mixed infection and therapy should be reviewed in such patients. A serological diagnosis of chlamydial infection is relatively easy and cheap and enables a rapid diagnosis of chlamydial infection to be made.
A case history of a 15-year-old schoolgirl with fluctuating bilateral uveitis, bilateral stromal keratitis with vascularisation, and bilateral deafness associated with tinnitus and balance disturbance is described. Three years from the onset of her clinical signs she died of a sudden cardiac arrest caused by endocarditis associated with valvular and arterial lesions. Chlamydia psittaci was isolated from her conjunctiva. In her blood type-specific antichlamydial antibody at a level of 1/64 against her own isolate was detected. The clinical findings in this patient were suggestive of a Cogan's syndrome. It is highly probable that the chlamydia isolated from the eyes was responsible for her various lesions.
A case of keratoconjunctivitis caused by adenovirus type 21 in London has been described. A 59-year-old woman attented hospital in August 1974 complaining of a 3-week history of redness, grittiness, watery discharge, and photophobia in her left eye and a slight upper respiratory infection. Clinical examination showed a moderate follicular conjuctivitis mainly in the lower and upper fornices, which lasted for 6 weeks. In the cornea a moderate amount of epithelial and subepithelial punctate keratitis was observed. The subepithelial opacities were coarse, discrete, and round and lasted for 4 months. The course of follicular conjunctivitis and the subepithelial punctate keratitis in this patient was similar to epidemic keratoconjunctivitis caused by adenovirus 8. A conjunctival swabbing collected from this patient was positive for adenovirus serotype 21.
During the 18 months January 1975 to June 1976, 25 cases of acute herpetic follicular conjunctivitis and keratoconjunctivitis resembling adenovirus ocular infection presented in the External Eye Disease Clinic, Moorfields Eye Hospital, City Road, London. Herpes simplex virus was isolated in HEp2 cells in 22 patients, and the remaining 3 patients were identified by a minimum 4-fold rise in the level of antiherpes simplex virus antibody in their blood. No adenovirus was isolated from these patients, but complement fixation test for adenovirus was positive in 1 patient with cultural test positive for herpes simplex virus. Most patients were between 20 and 35 years old and the ratio of males to females was 12 to 13. At the initial visit the clinical features of disease were moderate to severe conjunctival papillary and follicular reasons with epithelial and subepithelial punctate keratitis but little systemic disease. In the absence of typical herpetic lesions of face, lids, or cornea the disease resembled adenovirus types 8 or 19 keratoconjunctivitis. Of these 25 patients 5 subsequently developed typical herpetic lesions of lids or cornea. In the remaining 20 cases the correct diagnosis could be made only by cultural or serological tests. Virological diagnosis provides a rational basis for antiherpetic chemotherapy, which appears to shorten the course of infection.
A rapid serodiagnostic test for the diagnosis of paratrachoma (TRIC ophthalmia neonatorum, inclusion conjunctivitis, TRIC punctate keratoconjunctivitis, and trachoma of sexually transmitted origin) has been developed. The technique is based on using a modified micro-immunofluorescence test for detecting antichlamydial IgG and IgM in the blood and IgG and IgA in tears. The blood samples are collected on cellulose sponges after a finger prick, and tears are collected by introducing small sponges into the lower conjunctival fornix of the eye. The blood and tear samples collected in this way could be sent to the diagnostic laboratory by post without special arrangements for cold storage. In general the presence of antichlamydial IgG at a level of greater than or equal to 1/32 or IgM at a level of greater than or equal to 1/8 in blood and antichlamydial IgG or IgA at a level of greater than or equal to 1/8 in tears was closely associated with ocular paratrachoma. The combined results of the micro-IF test of blood and tears has yielded the highest rate of positivity (90%). In patients with acute untreated paratrachoma the sensitivity of this test was similar to that of irradiated McCoy cells. In patients with a milder infection receiving antibiotics the sensitivity of the serodiagnostic test was superior to that of the cultural test. The high sensitivity and specificity of this rapid, simple and inexpensive serodiagnostic test for the diagnosis of chlamydial ocular infections, coupled with simple and practical methods of collection and transport of blood and tear specimens, offer advantages over cultural tests for routine diagnosis and study of chlamydial ocular infections.
A predominance of TRIC serotype A has been isolated from schoolchildren in a population in Southern Tunisia with severe hyperendemic trachoma. The serotyping results correspond precisely with the serological findings in patients' tears and sera. Geometric mean titres of serum or tear antibody in defined populations or areas can thus give a useful indication of the prevalent serotypes. Collection of tear fluids on sponges is a more practical method than collection by filter paper strips and gives higher levels of antibody. The presence of antibody to Chlamydia trachomatis in tears correlates well with the presence of infectious agent in the eye and with the intensity of conjunctival inflammatory disease. The measurement of antichlamydial tear antibody can thus provide a meaningful index of the prevalence and intensity of active trachoma in a population. The role these antibodies may play in resistance to re-infection is not yet clear.
The sensitivities of Giemsa, immunofluorescence, and immunoperoxidase staining for the detection of Chlamydia psittaci inclusions in conjunctival scrapings and in irradiated McCoy cell monolayers were compared. Conjunctival specimens were obtained from a cat colony in which a trachoma-like disease, feline chlamydial keratoconjunctivitis, was endemic. The two immunochemical techniques were found to be of equal sensitivity and 50% to 100% more sensitive than Giemsa stain. Permanent preparations of immunoperoxidase stained material can be made and can be read using a simple light microscope. These features make the technique more useful than immunofluorescence staining, which gives temporary preparations that must be examined with a specialised fluorescence microscope.
The possible role of Chlamydia trachomatis in non-acute prostatitis was investigated by cultural and serological techniques in a study of 53 adult males. C. trachomatis was isolated from the urethra of only one of the 53 patients and from none of the 28 specimens of prostatic fluid from the same patients. By means of a modified microimmunofluorescent test, serum chlamydial IgG antibodies at a titre of 1/64 or greater, or IgM antibodies at a titre of 1/8 or greater, or both were detected in six of the patients, suggesting a recent or current chlamydial infection, while IgG or IgA antibodies at a titre of 1/8 or greater were detected in the specimens of prostatic fluid from two of the 28 men studied. In the seven patients with evidence of chlamydial infection, as well as in a further 13 of the 53 patients studied, the presenting symptoms suggested non-gonococcal urethritis (NGU) rather than prostatitis. Thus in this study C. trachomatis would appear to play a minor aetiological role, if any, in non-acute prostatitis.
A rapid serodiagnostic test for the presumptive diagnosis of chlamydial infection of the cervix has been developed. The method used in based on the modified micro-immunofluorescence test using pooled chlamydial antigens and the detection of different immunoglobulin classes of chlamydial antibody in sera and cervical secretions. The presence of IgG chlamydial antibody at a level of 1/64, or IgM antibody at a level of 1/8 or greater, or both in sera and IgG or IgA antibody at a level of 1/8 or more or both in cervical secretions was closely associated with the isolation of Chlamydia trachomatis and non-specific genital infection. In general, serodiagnosis was three to nine times more sensitive than cultural methods, and the detection of IgG chlamydial antibody in cervical secretions alone provided the most sensitive of the serological tests. This sensitive, low-cost, rapid, and simple serodiagnostic test for the presumptive diagnosis of chlamydial infection of the cervix, coupled with transportation of specimens by post, offers advantages over conventional isolation techniques for the routine diagnosis and management of chlamydial genital infections.
A new technique is presented using impression of the conjunctiva with a plastic device to study the conjunctival response in various conjunctival disease states. When used with a rapid acting stain the impression technique proved to be accurate, reproducible, and nondestructive when compared to the standard spatula scraping of the conjunctiva using Giemsa stain.