To determine:
The limitations of culture methods and PCR analysis in the diagnosis of pertussis.
Introduction:
Pertussis which is also called as whooping cough disease in the U.S. It is caused by Bordetella pertussis. It is a gram negative coccobacillus anaerobic bacterium that is highly transmissible by the respiratory droplets.

Explanation of Solution
There are some characteristics of Bordetella pertussis that enhanced the cases of pertussis infection during 2011 to 2012 in the Washington States. It was difficult to culture Bordetella pertussis, which were collected from nose and throat swabs. This bacterium is fastidious organism, which requires complex and specific nutrients to survive. Due to its fastidious requirements it was difficult to culture this bacterium.
The difficulty in culturing the specimens of Bordetella pertussis in a person who is received antibiotics is also the limitation for its culture method. Culture methods of Bordetella pertussis were time consuming due which there was delay in the treatment of suspected pertussis cases. Diagnosis of pertussis is also done by PCR, which is only sensitive in the first 3 weeks of pertussis cough. False negative or positive obtained through PCR are some of its limitation.
Fastidious nature of bacterium and false results of PCR are some of the limitations of Bordetella pertussis culture.
To determine:
The other diagnostic method that was used in diagnosing pertussis outbreak.
Introduction:
Person infected with whooping cough is characterized by paroxysmal or violent cough followed by abrupt intake of breath sounding like “whoop”. In the year 2012, there was an epidemic of pertussis in the Washington State.

Explanation of Solution
In the initial six months of the year 2012, around 2,520 pertussis cases occurred in Washington States with the most incidences seen in children between 10 to 14 ages. Bordetella pertussis is highly communicable, which is transmitted through airborne droplets or contact with infected mucous discharge. Due to limitations in culturing method and PCR for analysis of Bordetella pertussis some other tests were performed by the “Centers for Disease Control and Prevention” (CDC) in the outbreak of pertussis in Washington States.
CDC performed pulse-field gel electrophoresis (PFGE), which is a type of DNA fingerprinting to diagnose pertussis. PFGE is used to determine the larger fragment of DNA. In pulse-field gel electrophoresis, periodically
According to CDC around 54% of bacterial isolate represented the common isolate of Bordetella pertussis and about 20 isolates represented the seven less common bacterial isolates. About five samples were PFGE profiles had not been earlier observed in CDC database.
The other diagnostic method that was performed by CDC in diagnosing pertussis outbreak was pulse-field gel electrophoresis.
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Chapter 17 Solutions
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