This document outlines the diagnostic protocol for Neisseria gonorrhoeae, which causes gonorrhea. It details that N. gonorrhoeae is a gram-negative diplococcus that can infect warm, moist areas of the body. Symptoms usually occur within 2-14 days and include discharge and pain. Diagnosis involves culturing samples on selective media, conducting biochemical tests like catalase and sugar utilization, and using molecular methods like PCR. Treatment involves antibiotics like ceftriaxone and azithromycin.

1. DIAGNOSTIC PROTOCOL FOR
NEISSERIA GONORRHEA
-TEJESWINI.I
-B.Sc. Microbiology

2. Contents
▪ Introduction
▪ Causative agent
▪ Symptoms
▪ Infections caused
▪ Culture & identification
▪ Biochemical test & basic characteristics
▪ Molecular diagnosis
▪ Serological diagnosis
▪ Confirmatory test
▪ Drugs used in treatment

3. introduction
GONORRHEA IS A SEXUALLY TRANSMITTED
INFECTION (STI) .
WARM, MOIST AREAS OF BODY INCLUDING
URETHRA, EYES, VAGINA, ANUS, THROAT,
MALE AND FEMALE REPRODUCTIVE SYSTEM.

4. causative agent
▪ NEISSERIA GONORRHEA CAUSES GONORRHEA.
 GRAM NEGATIVE DIPLOCOCCI
 ISOLATED BY ALBERT NEISSER
 FASTIDIOUS ORGANISM
 OBLIGATE AEROBE
 SURVIVES WITHIN NEUTROPHILS

5. SYMPTOMS
▪ USUALLY OCCUR WITHIN 2-14 DAYS AFTER EXPOSURE.
▪ ASYMPTOMATIC CARRIER IS ALSO POSSIBLE
 IN MALES IN FEMALES
 URGENCY IN URINE DISCHARGE FROM VAGINA
 SWELLING IN TESTICLES PAIN IN LOWER ABDOMEN
 REDNESS IN PENIS SORE THROAT

6. SYMPTOMS IN MALE AND FEMALE

7. complications/infections CAUSED
 PELVIC INFLAMMATORY DISEASE (PID) IN FEMALES.
 EPIDIDYMITIS IN MALES
 OPTHALMIA NEONATORUM (CONJUCTIVITIS IN NEWBORN)

8. URETHRITIS

9. CULTURE & IDENTIFICATION
▪ NEISSERIA GONORRHEA IS
USUALLY ISOLATED ON THAYER -
MARTIN AGAR.
• THAYER- MARTIN AGAR
CONTAINS 5% CHOCOLATE SHEEP
BLOOD AND ANTIBIOTICS LIKE
VANCOMYSIN, COLISTIN ETC.

10. BIOCHEMICAL TESTS
▪ The SUPEROXOL TEST (catalase
test using 30% H2O2) was used
to differentiate Neisseria
gonorrhea from other Neisseria
species.
▪ A positive test was defined as
immediate, brisk bubbling upon
dropping 30% H2O2 onto a
bacterial colony.

11. Basic Characteristics
Properties ( Neisseria
gonorrhoeae)
Capsule Negative (-ve)
Catalase Positive (+ve)
Gas Negative (-ve)
Gram Staining Negative (-ve)
H2S Negative (-ve)
Hemolysis Negative (-ve)
Nitrate Reduction Negative (-ve)
Oxidase Positive (+ve)
Pigment Negative (-ve)
Shape Diplococci
DNase Negative (-ve)
Fructose Negative (-ve)
Glucose Positive (+ve)
Lactose Negative (-ve)
Maltose Negative (-ve)
Mannose Negative (-ve)
Sucrose Negative (-ve)

13. molecular diagnosis
▪ FDA( Food and Drug Administration) APPROVED
MOLECULAR METHODS ARE :-
 POLYMERASE CHAIN REACTION (PCR)
 TRANSCRIPTION-MEDIATED AMPLIFICATION (TMA)
 STRAND DISPLACEMENT AMPLIFICATION (SDA)

14. In this reaction, multiple copies of
the gene (or DNA) of interest is
synthesized invitro using two sets
of primers (small chemically
synthesized oligonucleotides that
are complementary to the regions
of DNA) and the enzyme DNA
polymerase.
POLYMERASE CHAIN
REACTION

15. SEROLOGICAL DIAGNOSIS
RAPID CARBOHYDRATE UTILISATION TEST
(RCUT)
In this test, patterns of acid production from the
carbohydrates – glucose, maltose, lactose, sucrose,
(and fructose) – are used to identify Neisseria and
related species. Production of acid from
carbohydrates is indicated by a change in the color of
the phenol red indicator from red to yellow.

16. serological diagnosis
 ENZYME LINKED IMMUNO SORBENT (ELISA)
 INDIRECT HAEMAGGLUTINATION (IHA)
 GONOCOCCAL COMPLEMENT-FIXATION TEST
(GCFT)

17. Confirmatory test
▪ THE RECOMMENDED METHOD IS NUCLEIC ACID
AMPLIFICATION TEST (NAAT).
▪ THIS TEST DETECTS THE GENETIC MATERIAL THAT IS
THE DNA OF NEISSERIA GONORRHEA
USUALLY ITS RECOMMENDED TO TAKE TEST
FOR OTHER STI’s ALSO AS THEY ARE ALL
CO-RELATED TO EACH OTHER.

18. DRUGS USED IN TREATMENT
CEFTRIAXONE
CEFOTAXIME
AZITHROMYCIN
ERYTHROMYCIN