Mycobacteria Part 3

MAHATHIR ALPHA · 55問 · 2年前

通報

問題一覧

Describe the appearance of Mycobacterium fortuitum in MAC

(+) growth on Mac Conkey agar without crystal violet

It mostly associated with disseminated cutaneous infections in immunocompromised patients.

Mycobacterium chelonae

It is also associated with infections of the skin, lungs and bone. It exhibits more resistance to antimicrobial agents than M. fortuitum.

Mycobacterium chelonae

Culture: rough or smooth nonpigmented colonies; does not produce extensive flamentous branching colonies.

Mycobacterium chelonae

What are the biochemical test and result of Mycobacterium chelonae?

(+) 3-day Arysulfatase test -Growth on Mac Conkey agar without crystal violet -Without nitrate reduction

What is the reservoir of Mycobacterium abscessus?

Tap water

What is the related infections and disease Mycobacterium abscessus?

Chronic lung disease Otitis media

What are the biochemical test and result of Mycobacterium abscessus?

(+) 3-day Arysulfatase test -Growth on Mac Conkey agar without crystal violet -Without nitrate reduction.

Microscopy: Long and tapered or short rods are curved, branching or y-shaped forms

Mycobacterium smegmatis Group

Culture: rough, wrinkled or coarsely folded smooth or rough with dense centers

Mycobacterium smegmatis Group

Describe the appearance of Mycobacterium smegmatis Group in MB 7H10 agar

Pigmentation is rare or late - nonpigmented to pink colonies

What species are included in the Mycobacterium smegmatis Group?

Mycobacterium smegmatis sensu strict Mycobacterium goodle Mycobacterium wolinskyi

What are the biochemical test and result of Mycobacterium smegmatis group?

(-) 3-day Arylsulfatase test (+) nitrate reduction and iron uptake (+) growth on Mac Conkey agar without CV and media containing 5% NaCI

Where should the procedures of Laboratory diagnosis of Mycobacteria be done?

Class II or III biological safety cabinet (BSC)

True/False: In the laboratory diagnosis of Mycobacteria, the area which the specimens and cultures are processed should not have negative air pressures in relation to other areas - the airflow should be from clean areas (corridors) into less clean areas (mycobacteriology lab).

False

True/False: In the laboratory diagnosis of Mycobacteria, all potentially infectious specimens should be tightly covered when they are outside the BSC.

True

What are specimens that can be used for the laboratory diagnosis of Mycobacteria?

Sputum Gastric lavage Urine Bronchoscopy specimen Fecal specimen Body fluids Blood Wound, skin lesion aspirates and tissue

What is the preferred sputum specimen for the laboratory diagnosis of mycobacteria?

Early morning sputum specimen

What is the required volume of sputum specimen?

5-10 mL aerosol-induced sputum

What is used for the collection of Gastric lavage?

Levine collection tube

For who do you use Gastric lavage in the laboratory diagnosis?

For senile, nonambulatory patients and children younger than 3 yrs old

What is the required volume of Gastric lavage specimen?

20-25 ml of gastric secretions

Explain the procedure for the collection of Gastric lavage

3 specimens should be collected within 3 days after overnight

Gastric lavage should be processed within _____ of collection or neutralized w/ ______________ to pH 7.0 - prolonged exposure to gastric acid kills mycobacteria

4 hrs sodium carbonate

What is the preferred urine specimen for the laboratory diagnosis of mycobacteria?

First morning midstream urine (3 consecutive days)

What is the required volume of Urine specimen in the laboratory disgnosis of Mycobacter?

15 ml or entire volume of voided urine

True/False: Urine may be collected through an indwelling catheter with a needle and syringe

True

It is the specimen of choice for detecting NTM

Bronchoscopy specimens

What are included in Bronchoscopy specimens?

Bronchoalveolar lavage (BAL) Bronchial washing Transbronchial biopsy

This specimen is used to identify patients who may be at risk of developing M. avium-complex disease, especially those with AIDS

Fecal specimens

True/False: Stool specimens should be submitted with a preservative

False

If processing of Fecal specimens will be delayed, the specimen should be frozed at ____ celsius

-20

What are included in the Body fluidsused in laboratory diagnosis of Mycobacteria?

Pleural fluid Pericardial fluid Peritoneal fluid Joint aspirate CSF

What should be used to collect Pleural, pericardial and peritoneal fluids?

Sterile anticoagulant (EDTA or heparin)

What is the required volume of Body fluids specimen?

CSF - 2 mL; for exudates and pericardial and synovial fluids; 10-15 mL for abdominal and chest fluids

This specimen is collected to diagnose individuals with AIDS

Blood

What is the culture medium used for blood in the laboratory diagnosis of mycobacteria?

BACTEC 12A vial

For wound and skin aspirates, the specimen are transferred in ___________.

Liquid medium

If the tissue specimens cannot be processed immediately, what should be added to prevent dehydration?

10-15 mL of sterile saline

These procedures are performed to kill all contaminating organisms (nonmycobacteria) and to dissolve mucous substances

Decontamination and Digestion

True/False: Dissolving (liquefying) the mucin prior to inoculation onto culture medium enables the mycobacteria to use the nutrients of the medium

True

True/False: The high lipid content in the cell wall of mycobacteria makes them more susceptible to the killing action of various chemical

False

What are the factors affecting the action of the decontaminating agent?

Concentration of the chemical agent Exposure/contact time Temperature

What are the specimens that require decontamination?

Sputum Voided urine Autopsy tissue Abdominal fluid Any contaminated fluid

What are the specimens that require decontamination and digestion?

Sputum Gastric washing Bronchoalveolar lavage (BAL) Bronchial washing Transtracheal aspirate

What are the specimens that does not require decontamination?

CSF Synovial fluid Biopsy tissue from deep organs

What are the reagents used in the Decontamination and digestion?

2-4% NaOH 5% Oxalic acid Zephiran-trisodium PO4 (Z-TSP) 1% Cetyl-Pyridium Chloride N-Acetyl-L-Cysteine (NALC)-NaOH

NACL is also known as what?

Dithothreitol

It both a decontamination and digestion agents.

N-Acetyl-L-Cysteine (NALC)-NaOH

It prolonged shelf life of sputum for 8 days (for transport)

1% Cetyl-Pyridium Chloride

It is a decontamination-digestion agent which are used for specimens containing large numbers of bacteria

Zephiran-trisodium PO4 (Z-TSP)

_____________- results in greater isolation of mycobacteria

Phosphate buffer

It is an effective decontaminant with little bactericidal effect on the tubercle bacilli

Zephiran

It liquifies sputum rapidly but requires a long exposure time to decontaminate the specimen

TSP

It is the most common decontamination agent. And it also serves as both a decontaminant and digestant.

2-4% NaOH

This reagent is used for decontamination of sputum specimens containing Gram-negative rods like Pseudomonas (cystic fibrosis).

5% Oxalic acid

Sputum specimens treated with this agent can be inoculated into broth culture

5% Oxalic acid

Mycobacteria possess cell walls that contain __________, which are long chain, multiple cross-linked fatty acid.

Mycolic acid

___________ is affected by age of colonies, medium for growth and exposure to UV light.

Acid fastness

True/False: Rapidly growing species appear to be acid fast-variable

True

If at least 2 of the first three sputum direct smears are positive, then 3 specimens are often sufficient to confirm a ___________.

Diagnosis

CSF, urine, and bone marrow specimes - concentrated specimens are used and heat-fixed for ________.

15 mins

Give out the Distinct acid-fast organisms

Mycobacterium Cryptosporidum Isospora

Give out the Partially acid-fast organisms

Nocardia Gordonia Rhodococcus Tsukamurella Legionella micdadel

What are the different AFB staining methods?

Zehl-Neelsen/ Hot stain procedure Kinyoun/Cold stain procedure Auramidine-Rhodamine

Also known as Hot stain procedure

Zehl-Neelsen

Also known as Cold stain procedure

Kinyoun

This AFB staining method is preferably for tissues

Kinyoun/Cold Stain Procedure

This AFB staining method is sensitive, reliable and specific

Auramine-Rhodamine

AFB are examined at what magnification?

250x 400x

Auramine is more sensitive than ___________.

Carbol fuchsin

True/False: Auramine-stained smears may be restained by the Ziehl-Neelsen or Kinyoun method.

True

What is the positive result for Auramine-Rhodamine?

Bright yellow-orange bacilli against a dark background

What are the characteristics of a False Negative Acid Fast Smear?

1. Overzealous decontamination 2. Loss from concentration techniques smear 3. Organisms obscured by a too thick 4. Over-decolorizing of the smear 5. Poor counterstaining 6. Lack of observer proficiency in reading stains

What are the characteristics of a False Positive Acid Fast Smear?

1. Changes in the cell wall 2. Insufficient decolorization 3. Laboratory contamination 4. Delayed processing 5. Overgrowth of other bacteria

What are two major type of culture media used in the laboratory diagnosis of Mycobacteria?

Solid media Liquid broth media

What are the media included under Solid media?

Egg based media Serum agar-based media

What are the media included under Liquid/Broth media?

BACTEC 12B (MB 7H12) BACTEC 13 B (MB 7H13)

In Solid media, culture are incubated at ______ in the dark with 5-10% CO2

35 celsius

What bacteria/s grows best at 30-32 degree celsius in Solid Media?

Mycobacterium marinum Mycobacterium haemophilum Mycobacterium ulcerans

What bacteria grows best at 42 degree celsius in Solid Media?

Mycobacteria xenopi

It is an inhibitory agent of nonmycobacteria

Malachite green

This media are composed of fresh whole eggs, potatoflour, glycerol, milk, potato and malachite green.

Egg-based media

What is the shelf-life of Egg based media?

One year

Give examples of Egg-based media

Lowenstein Jensen Petragnani medium Wallenstein medium

Most commonly used Egg-based media

Lowenstein Jensen

This Egg-based media is used for recovery of mycobacteria from heavily contaminatetd specimen

Petragnani medium

Petragnani medium contains ___________ of malachite green

0.052 g/dL

This egg-based media is used for M. avium complex

Wallenstein medium

It is also known as Transparent media

Serum agar-based media

This media is compossed of vitamins, salts, cofactors,glycerol, malachite green, agar, oleic acid, bovine albumin, glucose and beef catalase.

Serum agar-based media

Give examples of Serum Agar-based media

Middlebrook 7H10-11 Mitchison's medium 7H11

Excessive heating and light exposure of agar- based media result in the release of ___________________, which is toxic to mycobacteria

Formaldehyde

This serum agar-based media produce positive cultures in 3-4 weeks and contains 0.1% casein hydrolysate (MB7H11).

Middlebrook 7H10-11

This serum agar-based media contains polymyxin V, amphotericin B, carbenicillin and trimethoprim lactate

Mitchison's medium 7H11

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