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National Institute of Communicable Diseases |
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ANNUAL REPORT 1997 |
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Microbiology Division Microbiology
Division of the Institute is a composite unit consisting of well organised
sections of virology, bacteriology and mycology; each having specialised
objectives mentioned below: §
Referral diagnostic services for various infectious viz.,
viral hepatitis, polio, measles, coxsackie viruses, Rubella, Cytomegalo and
herpes viruses (RCH of TORCH gastroenteritis, tuberculosis and fungal
infections. §
Laboratory based dianostic
services and surveillance of the aforementioned
diseases. §
Laboratory
support to National Health Programmes. §
Imparting
training, organising workshops for
peripheral labhoratories for manpower development. §
Supply of diagnostic kits and
reagents. §
Storage and supply of vaccines. §
Supply of cell lines,
viral/bacterial/fungal cultures to medical colleges, teaching and research institutions. §
Compilation
and dissemination of laboratory manuals/teaching materials. §
Specialised
research and development activities. §
Help
in establishment of a full-fledged Biotechnology unit at NICD, primarily for
gene sequencing of wild and vaccine strains of polioviruses. §
Technical
support to National/International Health Agencies. Routine Services / Activities (Maintenance of stock cultures of
various animals, pathogens and parasites)
i.
Cholera/Enterobacteriaceae Laboratory
§
Laboratory
based surveillance of diarrhoeal diseases. §
Laboratory
support to outbreak investigation of diarrhoea. §
Referral
services for identification and serotyping of Vibrio cholerae and other enteric pathogens. §
Characterization
of various types of E.coli
ii.
ARI/Meningitis Laboratory §
Isolation
and characterisation of the bacterial etiological agents of acute Respiratory
infection. §
Serological
studies of Dipohtheria, Pertussis, Tetanus and Meningitis. §
Development
and characterisation of indigenous rapid diagnostic tests. §
Storage
and supply of meningococcal kits/ vaccines etc. §
Monitoring
of the proved cases of faecal diphtheria in collaboration with I.D.Hospital. §
Laboratory
support to outbreak investigations. §
Referral
Diagnostic services for characterisation of pathogens causing meningitis. iii. Polio/Enterovirus Laboratory §
Regional
reference Centre (WHO) for Polio diagnosis in South East Asia. §
Potency
testing of Oral Polio vaccine for cold chain monitoring. §
RNA
probe hybridisation and ELISA for intratypic differentiation of wild polio
viruses. §
Referral
services for Coxsackie B virus associated cardiomyopathies, SSPE. §
Diagnosis
and serological studies of Measles. §
Maintenance
of different cell lines for virus studies. §
Diagnostic
services for enteroviruses. iv. Hepatitis Laboratory §
Services,
research and training activities pertaining to viral hepatitis. §
Co-ordination
of National Viral Hepatitis Surveillance Programme. §
Regional
Centre for Viral Hepatitis Surveillance. §
Multicentric
study to ascertain aetiology of jaundice in community.
v.
Teratogenic Virus Laboratory §
Acts
as Centre for diagnostic support for rubella, CMV and Herpes simplex virus
infections in cases of pregnant women in and around Delhi. §
Diagnosis
of viral aetiology of congenial malformations in newborn babies. vi. Tuberculosis Laboratory §
Maintaining
the cultures of mycobacteria isolated from various clinical samples from
suspected cases of tuberculosis. §
Serodiagnosis
of tuberculosis. §
Antimycobacterial
drug sensitivity testing. vii. Medical Mycology Laboratory §
Processing
of clinical samples by direct microscopy, culture, serology, identification of
fungal pathogens, preservation of cultures, providing diagnostic services on
mycosis on request from Hospital/Institutions. viii.
Environmental Microbiology
Laboratory §
Bacteriological
analysis of water samples. §
Evaluation
of field kits for water testing. §
RNA
probe hybridisation and ELISA for intratypic differentiation of wild polio
viruses. §
Referral
services for Coxsackie B virus associated cardiomyopathies, SSPE. §
Diagnosis
and serological studies of Measles. §
Maintenance
of different cell lines for virus studies. Maintenance of live material/cell cultures Cell Lines:
i.
Hep2
Cincinnati cell line
ii.
HELA
iii.
Vero
iv.
RD
(Rhabdomyosarcoma)
v.
L20B Live Materials:
i.
More
than 100 isolates of various fungal cultures of medical importance
ii.
Virus
maintained are:
iii.
Polio
type 1, 2, 3 (Sabin and Wild)
iv.
Coxsackie
B1-B6
v.
Some
non-polio Enteroviruses Bacterial cultures:
i.
Some
non-polio Enteroviruses
ii.
Vibrio
cholerae
iii.
C.diphtheriae
iv.
Mycobacterial
cultures
v.
Other
enteropathogens Supply of live material §
On
request 5 different fungal cultures viz. Aspergillus
flavus, Sporothrix schenckii, §
Madurella grisea, Trichophyton
mentagrophytes and Cladrosposirum spp. were supplied to Shimla Medical College. Similarly 4
different fungal cultures viz. §
Cryptococcus neoformans,
Sporothrix schenckii, Dermatophyte and Aspergillus flavus were supplied to Deptt. of
Microbiology, B.J. medical College, Civil Hospital, Ahmedabad (Gujarat). §
Cell
cultures HEP2, RD cell line vero cell line are frequently supplied to all polio
national laboratories and OPV testing laboratories from time to time. §
Supply
of Vibrio cholerae and other enteric
pathogens isolates for workshops, training and teaching purpose to the
different part of country as demanded. §
V.cholera. 01,V.cholerae
0139,Salmonella sp. Shigella sp. and NCTC E.coli. §
Clinical
specimens for quality control purpose. §
Loeffler’s
serum slopes to I.D. Hospital, Delhi. §
Cary
Blair Transport Medium to I.D. Hospital, Delhi and other Institutes. §
Teaching
slides for viral, bacterial and fungal pathogens. §
Virological §
MEM,
Hanks BSS, Enteroviruses typing antisera. §
Fungal §
Serological
reagents produced by the section for diagnosis of Aspergillosis. Storage of reagents/materials §
Meningitis
vaccine: 71050 doses supplied to 25 states and Uts for HAJ pilgrims. §
FCS
and Microtitre plates to Polio Vaccine Testing Labs in the country. §
Hepatitis
kits to different surveillance centres in the country. §
Bacterial,
viral and fungal typing antisera for typing and identification purposes. Diagnostic support services The
division provides diagnostic support services in various diseases: Viral Hepatitis
Virus isolation from AFP Cases Polio
1 – 124
Wild (cases 520) Polio
2 – 6
(3 Sabin) Polio
3 – 21
(5 Wild, 1 Sabin) Non-Polio
enterovirus – 6 (3 Sabin) Mixture
of Polio – 28 Tuberculosis
Enteropathogens
Medical mycology
Public health and other biological services Bacteriological
analysis of drinking water provided to the following beneficiaries:
i.
Airport
Health Authority – in connection with VVIP flights going abroad; on request
ii.
Govt.
hospitals and other institutions; on request
iii.
General
public: by reference. Identification services §
Identification
of V.cholerae and other
enteropathogens (Salmonella, Shigella and Yersinia) §
Identification
of different mycobacterial isolates sent from other laboratories by carrying
out different biochemical tests. §
Identification
of Polio isolates obtained from different polio laboratories in the country. §
Identification
of fungal isolates obtained from different hospitals in Delhi. Serological tests §
ELISA
test for Tuberculosis. §
Immunodiffusion
and Latex agglutination test for fungal infections. §
Neutralization
test for Coxsackie B virus, Measles and Polio virus antibodies. §
Haemagglutination
Inhibition test for Measles. §
ELISA
test for various markers of Viral hepatitis A, B, C, D and E. §
ELISA
test for antibodies against different teratogenic viruses. §
Latex
agglutination test for meningitis (Antigen detection in CSF). Research Projects 1. Laboratory diagnosis and
surveillance of Cholera in Delhi In the
year 1997, 2532 rectal swabs were tested from the cases of gastroenteritis from
I.D. Hospital, Delhi and 72 from area in and around Delhi for the isolation of V.cholerae. Of these specimens, 937
(37%) were positive for V.cholerae 01
Ogawa serotype, 22 were V.cholerae 0139 (0.87%) and 54 were NGA vibrios (2.13%)
from I.D. Hospital, Delhi. Of the 72 cases from in and aground Delhi, 2 cases
were positive for V.cholerae 01 and
one was NAG vibrios. The incidence of V.cholerae
0139 was higher as 22 cases were identified in compare with previous year. On
serotyping, all the V.cholerae 01
isolates were found to be Ogawa serotype as previous years. Strains of V.cholerae were received from different
places for identification, serotyping and biotyping are given below:
2. Studies on gastroenteritis at
Delhi council for Child Welfare, Delhi (DCCW) 168
stool specimens were tested for enteropathogen collected from cases of
gastroenteritis from Council for Child Welfare, Delhi. Of these specimens, 15
specimens yielded Each coli pure culture, 13 Salmonella sps. None of cases were positive for Shigella. Of these 13 Salmonella,
eight were Salmonella factor (8) and
five were Sal. Factor (4,5). IOI stood specimens were also tested from the
staff members of Delhi Council for Child Welfare (DCCW), Delhi to assess the
carrier of enteropathogens i.e. Salmonella and Shigella among the staff. Of
these specimens, 3 yielded Salmonella,
one Shigella, 8 Esch.coli pure culture were also detected 160 stool
samples were tested for Rota-virus by Latex Agglutination test method in
infacts from Delhi Council for Child Welfare, Delhi. Of these specimens, 16
cases yielded Rota-virus positivity. 3. Studies on enteropathogens other
than V.cholerae The
project was to look for other important enteropathogenis i.e. Salmonella, Shigella, Esch.coli pure
cultures, Yersinia etc. in the cases of gastroenteritis. Of the 2604 rectal
swabs process, 146 yielded Esch.coli
pure culture, 25 Salmonella species,
19 Shigella species. For species
identification of Salmonella, the
isolates were sent to C.R.I., Kasauli (H.P.). Out of 19 Shigella species, Shigella
flexneri was predominant (8), followed by Shigella boydii (6). Shigella
dysenteriae (3) and Shigella sonnei
(2). E.coli serogrouping – out of 121
E.coli isolates sent to CRI, Kasauli
for serogrouping. 9 were founded to being to Enterotoxigenic (ETEC), 5 to
Enteropathogenic (EPEC), 2 to Enteroinvasive (EIEC) and 1 to Enterohaemorrhagic
category of diarrhoegenic E.coli. All
the pathogenic isolates were sent to antibiotic/drug resistance monitoring
laboratory for antibiograms. 4. Monitoring of faecal diphtheria in
and around Delhi 185
throat swabs from suspected diphtheria cases were received from I.D. Hospital,
Delhi, of these 19 were positive for C.diphtheriae.
All the isolates were sensitive to Penicillin, Erythormycin, and other commonly
used antibiotics. 5. National monitoring of cold chain
system: potency testing of field samples of OPV This is
an on-going activity with the aim to monitor cold chain system in the country.
This project is in collaboration with Ministry of Health & F.W. and
supports polio eradication activity. States which send the OPV field samples
are –U.P., Delhi, Rajasthan, Bihar and Madhya Pradesh. Potency
testing of field samples of OPV, 1997
During
1997, a total of 3640 samples of OPV samples from different states were tested.
All the samples were subjected to total virus concentration by microtitration
technique using Hep2 cell line. The results are shown in Table above. A total
of 73.20% samples were found to be having a satisfactory titres indicating that
cold chain condition is good but needs more strengthening at the peripheral
level. 6. Laboratory based surveillance of
Acute Flaccid Paralysis in Delhi and surrounding areas This is
an on-going project under the polio eradication activity of the country. During
1997, a total of 520 cases of AFP were reported to polio laboratory and 819
stool samples were collected from these cases. Two stool samples with 14 days
of onset of paralysis were collected only in 394 cases. As compared to previous
year, collection of samples has improved though still we still need to
strengthen it. All the samples after collection and pre-treatment were
subjected to polio virus isolation in RD and Hep2 cell line. Acute
Flaccid Paralysis cases – 1997
Intratypic
differentiation of polio isolates from AFP cases
The
virus isolates were subjected to polio and enterovirus typing. Results are
shown in the table. Fifty two samples showed the presence of non-polio
enterovirus. All the polio isolates
were subjected to intratypic differentiation by ELISA and probe
technique. All were found to be wild type except two polio type 2 isolates. 7. Epidemiology of Subacute Sclerosing Pan Encephalitis
(SSPE) in Delhi and surroundings Of the
important late sequalae of measles virus infection is SSPE. From all the major
hospitals of Delhi, a total of 963 clinically suspected cases of SSPE were
reported to the laboratory. These cases have evidence of SSPE by Cat Scan and
EEG findings. Serum and CSF samples were collected from each case to confirm
the diagnosis. Both
samples were subjected to presence of antibody titre by HAI test using monkey
erythrocyutes. Forty cases were confirmed to be having SSPE, as high antibody
titres were present in serum and CSF. These
positive cases were in the age group of 4 years to 18 years and predominantly
in male population. So far no case have been reported following measles
vaccination. The polio isolates from national laboratory at Bombay,Madras,
Ahmedabad and Kasauli were referred to regional laboratory at NICD for intratypic
differentiation. No polio type 2 wild virus was seen in 1997. 8. Role of RUBELLA, CMV and HSV2 in
congenital defects and abortions A total
of 557 samples were received from women and malformed babies belonging to
different clinical groups, attending Gynae OPD a various hospitals of Delhi and
surrounding areas. These samples were subjected to ELISA test for Rubella, CMV
and HSV-2 to show the presence of IgM/IgG antibodies against the above
infections. The detail result is shown in the table. Prevalence
of CMV, HSV and Rubella infection in BOH and congenitally malformed babies
9. Prevalence of various hepatitis
markers in acute and chronic viral hepatitis in and around Delhi Acute
sporadic jaundice cases/chronic active hepatitis/cirrhosis/portalhypertension/
chronic renal failure/haemodialysis patients, etc. have been studied for
hepatitis markers for hepatitis A, B, C, D and E in the serum samples. The
detail results are shown in the table. Prevalence
of hepatitis markers in acute and chronic viral hepatitis in and around Delhi
Earlier.
HBsAg was found positive in 243 out of 1352 cases screened while acute activity
out of those positive was seen in only 100 cases, while hepatitis C prevalence
specially among high risk individuals was seen in 18/103 cases. 10. Evaluation of various diagnostic
modalities in smear negative pulmonary tuberculosis, and role of estimation of
anti A60 and anti 38 kd antibodies in the diagnosis of cutaneous tuberculosis So far
15 serum and bronchial aspirates samples have been subjected to ELISA test; of
these 9 serum samples and 7 bronchial aspirates are positive for anti A60
antibodies. So far twenty-five serum samples obtained from suspected cases of
skin tuberculin. 50% were found positive for anti 38kd antibodies and 55%
positive for Anti A60 antibodies. 11. To monitor the drug resistance
pattern in clinical drug resistant/treatment failure and relapsed cases of
pulmonary TB So far
65 samples have been tested against INH, Streptomycin (SM), Ethambutol,
Rifampicin, Thiacetazone and Pyrazinamide. Results of 45 isolates are
available, and out of these, 42.55% are resistant to INH, 55.5%l to SM, 47.5%
to Rifampicin and 16.6% to Ethambutol. No resistance was seen towards
Thiacetazone and Pyrazinamide. 12. Water quality in various
environments in and around Delhi The
water samples collected from different environments – rural, semi-urban and
urban in and around Delhi were tested for bacteriological standards by the
conventional multiple tube method and
the Ejkman’s test. The water samples tested consisted of running and stored tap
water, running and stored hand pump water, well water, mineral water bottles.
From an analysis of the test results obtained, the following interesting
observations can be made: Bacteriological
Testing of drinking water samples
ND = Test not done due to insufficient
sample; RTW = Running tap water, STW = Stored tap
water; MBW = Mineral Water Bottle; RHPW = Running hand pump water; SHPW = Stored hand pump water; SPW = Swimming
Pool Water In the
first category, samples from flight kitchens of airline caters which has ideal
conditions of urban hygiene and water supply, only 75% of the samples tested
proved satisfactory (“Excellent”). The remaining 25% of unsatisfactory samples
included a large number of mineral water bottles. This needs to be further
confirmed and the causes for the same be investigated, since the mineral water
bottles are gaining popularity and wide usage, and hence might give a false
sense of security to consumers. 50% of
the water samples collected during the investigation of cholera and
gastroenteritis outbreaks were found to be unsatisfactory. Large majority of
these samples were from semi-urban and rural areas and from economically lower
starta. It is also observed that the samples were from stored hand pump and
stored tap water. Even
prominent hospitals and offices in the city yielded unsatisfactory water
samples, indicating poor maintenance of supply lines or storage conditions.
Accordingly, remedial measures were suggested. 13. Reliability checking of H2S-Strip
vial water test kits In
recent years a simple and easy to perform (even by untrained personnel)
“Yes-No” test for bacteriological quality of water has been devised. This
method, which is called “H2S-Strip” method, is currently under field evaluation
and quality standardization. It is expected to be adopted as the field test for
water quality monitoring in the hands of peripheral health workers and
community participants. Three batches of Commercially manufactured H2S-Strip
vial kits from two manufacturers were referred by the UNICEF, Delhi, for
reliability checking. The test kits were tested for false positivity, false
negativity and sterility standards. While the kits from one manufacturer showed
lot of false positivity, those from the other manufacturer showed false
negativity (much delayed positivity). The sterility standards of all vials
tested were, however, satisfactory. The results were sent to UNICEF for further
action at their end. 14. Studies on Candidiasis and Candida
spp. involved in patients of AIDS Candidiasis
is one of the common opportunistic infections in HIV infected persons.
Oropharyngeal, oesophageal and vulvo vaginal candidiasis are the most prominent
ones. However, the frequency of incidence of this disease in Indian AIDS
patients is not clearly understood. Also, although candiadiasis is known to be
caused mainly by Candida albicans, other specifies also have been encountered
to cause this disease. Therefore, a study was undertaken to find out the
frequency of this disease and the specifies involved in AIDS patients in
collaboration with Dr. Ram Manohar Lohia Hospital, New Delhi and the National
AIDS Control Organization. During the year under report, 16 AIDS patients have
been investigated and out of which, I were found culture positive for oral
candidiasis. Out of 9 positive cases 8 were positive for Candida albicans and
the identity of one is yet to be established. The study is in progress. 15. Studies on the frequency of
Opportunistic fungal infections in Kidney transplant patients Owing
to long duration of immunosuppressive therapy, an increasing number of patients
with organ transplantations with T & B cell defects are prone to serious
opportunistic infections. Since there is no detailed study on the frequency of
various fungal infections in organ transplant recipient in India, the present
study is initiated during the year in collaboration with the
Nephrology/Transplantation Unit of Batra Hospital and Medical Research Centre,
New Delhi. So far 9 transplanted patients were investigated and 2 were found
positive for systemic Candidiasis and one to Aspergillosis. The study is in
progress. 16. Studies on the incidence of fungal
meningitis in Delhi In view
of the sharp increase in the incidence of fungal meningitis in
immunocompromised patients in recent years, especially in patients of AIDS, a
study was underway to find out the magnitude of this problem in Delhi. Out of
27 patients investigated this year, 3 were found positive for Cryptococcosis
and one case was positive for Candidiasis. In the past, meningitis due to fungi
other than Cryptococcus was rarely known, but in recent years, it is being
frequently diagnosed in many parts of the world. 17. Incidence of pulmonary/systemic
mycosis in Delhi In an
on-going diagnostic survey conducted to find out the incidence of pulmonary/
systemic mycosis in patients of various chest and other disorders, it was found
that out of 49 patients investigated this year, 16 cases were found positive
for systemic Candidiasis and 5 were positive for Aspergillosis. It is
interesting to note that most of the cases diagnosed are from
immuno-compromised patients. 18. Studies on the incidence of
subcutaneous/cutaneous mycosis In a
collaborative study with various hospitals on the extent of prevalence and
etiology of subcutaneous mycosis, out of 9 cases investigated, 4 cases of
mycetomas were detected, (2 cases were due to Actinomycotic mycetoma, 2 cases was due to fungal mycetomas); one
case was due to sporotrichosis. Regarding cutaneous infections, one case was
diagnosed during the year and it was due to Trichophyton
rubrum. 19. Studies on the distribution and
epidemiological significance of the serotypes of Cryptococcus neoformans Studies on the isolation of the strains of C.neoformans from clinical as well as environmental sources were continued. 3 strains of C.neoformans were isolated during the year and were added to the poor of isolates preserved for serotyping. Six isolates have been confirmed positive for C.neoformans var. gath out of the list and the rest are under investigation for confirmation. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
