Laboratory diagnosis of pertussis infections: the role of PCR and serology.
This study reports on practical laboratory aspects of pertussis diagnosis. PCR assays were applied to respiratory specimens obtained during a large study of infants (less than 5 months old) admitted to paediatric intensive care units (n = 122), children (less than 15 years old) admitted to paediatric wards (n = 16) and their household contacts (n = 320). Estimation of antibodies to pertussis toxin and culture for Bordetella pertussis were attempted on specimens from the same patients, where available, and the overall utility of the diagnostic PCR assays was assessed by comparison to these results. A PCR assay for the human mitochondrial cytochrome oxidase (HMCO) gene was used for quality control of the extracted samples and an internal process control (IPC) was included in each sample to test for PCR inhibition. Four of 458 samples were considered unsuitable (three HMCO negative, one IPC negative) and excluded from further analyses. Positive PCR results were considered valid if they were either (i) positive for both of two B. pertussis gene targets (pertussis toxin S1 promoter and the insertion element IS481), i.e. consensus PCR positive, or (ii) repeatably positive in only one assay. Using these criteria, 52 of 454 (11.5 %) samples were considered as PCR positive for B. pertussis. Six of 356 samples were culture-positive for B. pertussis, 1/88 infants, 3/14 children and 2/254 contacts, giving an overall isolation rate of 1.7 %. Using these data, PCR gave an almost fivefold increase in diagnostic yield compared with culture (McNemar's test; P < 0.0001). Sera from 9/111 infants, 5/10 children and 14/210 contacts were positive. Serology and PCR results showed a high level of agreement (113/121) for infants and children. PCR demonstrated a significant improvement in diagnostic yield over culture. Serological testing also resulted in a significant increase in diagnostic yield compared to culture alone. PCR is a useful technique, but validity of results must be assured by careful control. Rapid diagnosis of B. pertussis infection particularly in infants by PCR, together with serological assays, can enhance surveillance systems for pertussis in all age groups.
Fry
Norman K
NK
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Tzivra
Oceanis
O
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Li
Y Ting
YT
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
McNiff
Anthony
A
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Doshi
Nivedita
N
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Maple
P A Christopher
PAC
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Crowcroft
Natasha S
NS
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Miller
Elizabeth
E
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
George
Robert C
RC
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
Harrison
Timothy G
TG
Respiratory and Systemic Infection Laboratory1 and Special Projects Laboratory2, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK.
D000293:Adolescent; D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood*; D001886:Bordetella pertussis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002648:Child; D004269:DNA, Bacterial / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007223:Infant; D007362:Intensive Care Units; D009305:Nasopharynx / Q000378:metabolism / Q000382:microbiology; D037342:Pertussis Toxin / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011159:Population Surveillance; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D014132:Trachea / Q000378:metabolism / Q000382:microbiology; D014917:Whooping Cough / Q000097:blood / Q000175:diagnosis*
D000907:Antibodies, Bacterial; D004269:DNA, Bacterial; C083388:pertussis toxin, S1 subunit; D037342:Pertussis Toxin