Journal of Postgraduate Medicine
 Open access journal indexed with Index Medicus & ISI's SCI  
Users online: 1144  
Home | Subscribe | Feedback | Login 
About Latest Articles Back-Issues Articlesmenu-bullet Search Instructions Online Submission Subscribe Etcetera Contact
 :: Next article
 :: Previous article 
 :: Table of Contents
 ::  Similar in PUBMED
 ::  Search Pubmed for
 ::  Search in Google Scholar for
 ::Related articles
 ::  Article in PDF (21 KB)
 ::  Citation Manager
 ::  Access Statistics
 ::  Reader Comments
 ::  Email Alert *
 ::  Add to My List *
* Registration required (free) 

  IN THIS Article
 ::  Abstract
 ::  Introduction
 ::  Methods
 ::  Results
 ::  Discussion
 ::  Acknowledgment
 ::  References
 ::  Article Figures
 ::  Article Tables

 Article Access Statistics
    PDF Downloaded103    
    Comments [Add]    
    Cited by others 3    

Recommend this journal


Year : 1992  |  Volume : 38  |  Issue : 2  |  Page : 72-4

Detection of serum antitrichomonal antibodies in urogenital trichomoniasis by immunofluorescence.

Dept of Microbiology, LTMM College, Sion, Bombay.

Correspondence Address:
R Bhatt
Dept of Microbiology, LTMM College, Sion, Bombay.

Login to access the Email id

Source of Support: None, Conflict of Interest: None

PMID: 0001432833

Rights and PermissionsRights and Permissions

 :: Abstract 

Trichomonas vaginalis is a frequently encountered genital pathogen in both males and females. In females, vaginitis due to this parasite is one of the most common manifestation. The indirect fluorescent technique (IFA) test was carried out to detect antitrichomonal antibodies in 370 female patients using whole cell antigen. Seventy one (19.18%) gave positive reaction for either of the class IgG, IgM and IgA antibodies. The level of the IgG class antibodies was found to be higher i.e. 58 (81.69%) than IgM 11 (15.27%) antibodies, which may be suggestive of past infection or a prolonged manifestation by the organisms.

Keywords: Antibodies, Anti-Idiotypic, blood,Evaluation Studies, Female, Fluorescent Antibody Technique, standards,Hospitals, University, Human, Immunoglobulin A, immunology,Immunoglobulin G, immunology,Immunoglobulin M, immunology,India, epidemiology,Mass Screening, Outpatient Clinics, Hospital, Sensitivity and Specificity, Trichomonas Vaginitis, blood,epidemiology,immunology,

How to cite this article:
Bhatt R, Pandit D, Deodhar L. Detection of serum antitrichomonal antibodies in urogenital trichomoniasis by immunofluorescence. J Postgrad Med 1992;38:72

How to cite this URL:
Bhatt R, Pandit D, Deodhar L. Detection of serum antitrichomonal antibodies in urogenital trichomoniasis by immunofluorescence. J Postgrad Med [serial online] 1992 [cited 2023 Jun 6];38:72. Available from:

  ::   Introduction Top

Trichomonas vaginalis, a flagellated, frequently encountered genital pathogen is responsible for trichomoniasis in both males and females. In women, the characteristic of trichomonial vaginitis, is a thin to a green frothy discharge.

Besides widely used microscopy and cultural technique to detect T Vaginalis, detection of antibodies to the parasite may help in diagnosis and detection of the asymptomatic carrier or a person with a past history of a trichomonas infection. Immunologic investigation of T vaginalis did not come into the picture until the mid 1940's, when the use of antibiotics allowed the establishment of axenic cultures of this species[1].

The present study was carried out to detect the antitrichomonal antibody level in serum, from female patients by an indirect fluorescent antibody (IFA) technique[2].

  ::   Methods Top

Three hundred and seventy female patients attending the gynecological outpatients department, with complaints of vaginal discharge, burning micturition and dysuria, were screened for the presence of Tvaginalis and circulating antibodies in their sera. T vaginalis was isolated and maintained in All Culture (AC) medium[3] supplemented with 10% inactivated horse serum, penicillin G (1000 ug/ml) and amphotericin B (250 ug/ml). The cultures were maintained axenically by subculturing every 48 - 72 hrs.

Preparation of Antigen:

T vaginalis cells in an exponential growth phase (24 hrs culture) were washed twice with 0.01 M phosphate buffer solution (PBS) (pH 7.2). The pellet was suspended in 10% formalin in PBS for 10 min. The cells were washed twice with PBS and resuspended in PBS to get 1 x l06 organisms per ml and stored at-20?C.

Control sera

Pooled sera from patients from whom T vaginalis was isolated represented positive control sera, whereas 100 sera obtained from children below 8 yrs age group represented negative control sera.


Standard indirect fluorescent antibody (IFA) technique was used. A drop of formalin fixed antigen was allowed to air dry in 0.5mm diameter wells on a glass slide. A serial two fold dilution of sera ranging from 1:2 to 1:64 was made in PBS and was layered on antigen coated wells which was allowed to react for 30 min at 37?C followed by three washings, each lasting for 15 min. The slides were air dried and reincubated for 30 min at 37?C with antihuman fluorescein labelled lgG, lgM and lgA (Behring) at 1/16 dilution. Slides were again washed three times as described previously and air dried. Finally, a drop of mounting fluid (9 parts glycerol + 1 part PBS pH 7.2) was layered and the preparation was examined under fluorescent microscope with high pressure HBO 200 mercury lamp.

  ::   Results Top

The positive slides when examined under fluorescent microscope, showed bright apple green fluorescence and the reaction was considered positive when at least 50% of the cells showed bright fluorescence [Figure - 1]. Out of 370 sera screened, antitrichomonal antibodies were detected in 71 cases (19.18%), whereas the parasites were isolated in 51 (13.78%) cases and smear was positive in 45 (12.16%)cases. All the control sera showed negative results [Figure - 2]. Of 71 (19.18%) cases positive by IFA, 58 (81.69%) were positive for IgG antibodies, whereas IgM class antibodies and lgA class antibodies were found in 11 (15.27%) and 2 (2.77%) cases respectively. [Table - 1].

Of 71 IFA positive cases, 50 (70.42%) cases correlated with culture technique, whereas 21 cases failed to show any correlation. these 21 cases were culture negative but IFA positive. Only one case of the 370 was I FA negative but culture positive [Table - 2].

  ::   Discussion Top

McEntegart et al4 in 1952 identified T vaginalis by the direct immunofluorescence reaction and succeeded in differentiating T vaginalis from T foetus.

Of 370 patients screened, 71 (19.18%) gave positive fluorescence for one of the antibody class, whereas culture was recorded positive in 51 (13.78%) cases only. Correlation of 70.42% was observed between culture and IFA techniques. Of 51 (13.78%) culture positive cases, 50 (98.03%) correlated with IFA technique, whereas only one case showed culture positive but IFA negative results, which is in close accordance with Kramar and Kucera 2, who reported 100% correlation between both the techniques.

Of total 71 positive fluorescence cases, 81.69% of cases were positive for IgG class antibodies whereas IgM and lgA class antibodies were observed in 15.27% and 2.77% of cases respectively. Detection of high titre IgG antibodies ( ? 1:16) among 65% of culture positive cases suggests either active or prolonged infection with T vaginalis, where as failure to detect high titre of IgG antibodies in culture negative cases suggests past infection. Ackers et al5 and Su KE6 have also reported the presence of IgG antibodies in serum because of systemic response to released antigen.

  ::   Acknowledgment Top

The authors wish to thank Hoechst (Behring), for providing antihuman fluorescein labelled antibodies for the present work.

 :: References Top

1. Honigberg BM. In: Kreier JP, Ed Parasitic Protozoa Vol. 11, Academic Press, Inc; 1978, pp 275-454.  Back to cited text no. 1    
2.Kramar J, Kucera K. Immunofluorescence demonstration of antibodies in urogenital trichomoniasis. J Hyg Epidemiol Microbiol Immunol 1966; 10:85-88.  Back to cited text no. 2    
3.Chatterjee DK, Ray DK. A simple medium for in vitro culture of Trichomonas vaginalis J Parasitol 1979; 65:815-816.  Back to cited text no. 3    
4.McEntegart MG. The application of haemagglutination technique to the study of Trichomonas vaginalis infection. J Clin Pathol 1952; 5:275-280.  Back to cited text no. 4    
5.Ackers JP, Lumdsen RD, Catterall RD, Coyle R. Antitrichomonal antibody in the vaginal secretions of women infected with Trichomonas vaginalis. Br J Vener Dis 1975; 51:319-323.  Back to cited text no. 5    
6.Su KE. Antibody to Trichomonas vaginalis in human cervicovaginal secretion. Infect Immun 1982; 37:852-857.   Back to cited text no. 6    


[Figure - 1], [Figure - 2]


[Table - 1], [Table - 2]

This article has been cited by
1 Not so benign: the clinical manifestations, diagnosis and treatment of trichomoniasis in men
Tucci V, Toney JF
ASIAN BIOMEDICINE. 2008; 2(2): 91-100
2 Plasma antibodies against Trichomonas vaginalis and subsequent risk of prostate cancer
Sutcliffe S, Giovannucci E, Alderete JF, et al.
3 A meta-analysis of the Papanicolaou smear and wet mount for the diagnosis of vaginal trichomoniasis
Wiese W, Patel SR, Patel SC, et al.
AMERICAN JOURNAL OF MEDICINE. 2000; 108 (4): 301-308


Print this article  Email this article
Previous article Next article
Online since 12th February '04
2004 - Journal of Postgraduate Medicine
Official Publication of the Staff Society of the Seth GS Medical College and KEM Hospital, Mumbai, India
Published by Wolters Kluwer - Medknow