The use of nested polymerase chain reaction and restriction fragment length polymorphism for the detection and typing of mucosal human papillomaviruses in samples containing low copy numbers of viral DNA

2002 ◽  
Vol 105 (1) ◽  
pp. 159-170 ◽  
Author(s):  
Patti Kay ◽  
Kathleen Meehan ◽  
Anna-Lise Williamson
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Fragment Length Polymorphism ◽  
Human Papillomaviruses ◽  
Chain Reaction ◽  
Viral Dna ◽  
Nested Polymerase Chain Reaction ◽  
Copy Numbers ◽  
Polymerase Chain ◽  
Restriction Fragment Length

scholarly journals Genotyping of Cryptosporidium species in children suffering from diarrhea in Sharkyia Governorate, Egypt

2021 ◽  
Vol 15 (10) ◽  
pp. 1539-1546
Author(s):  
Samira Metwally Mohammad ◽  
Magda Saad Ali ◽  
Sara Ahmed Abdel-Rahman ◽  
Raghda Abdelrahman Moustafa ◽  
Mohamed Hassan Sarhan
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Cryptosporidium Species ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Introduction: The protozoan parasite Cryptosporidium is one of the principal reasons for childhood diarrhea around the world. This work aimed to differentiate Cryptosporidium species among children suffering from diarrhea in Sharkyia Governorate, Egypt. Methodology: A total of 97 fecal specimens were taken from children suffering from diarrhea, attending Pediatric Clinics of Zagazig University and Al-Ahrar Hospitals. Full history was taken. Stool samples were examined microscopically using modified Ziehl–Neelsen stain for detection of Cryptosporidium oocysts. To identify Cryptosporidium genotypes, positive samples were then subjected to nested Polymerase chain reaction-restriction fragment length polymorphism targeting Cryptosporidium oocyst wall protein gene. Results: The overall detection rate was 27.8% (27/97) using modified Ziehl–Neelsen stain staining method. Using nested polymerase chain reaction, the gene was amplified in 85.2% (23/27). Restriction fragment length polymorphism analysis revealed that 65.2% (15/23) were Cryptosporidium hominis, 30.4% (7/23) were Cryptosporidium parvum, and one sample was not typed (4.4%). The significant risk factors associated with Cryptosporidium infection in children were animal contact and residence in rural areas. Conclusions: Cryptosporidium is a common enteric parasite affecting children in Sharkyia Governorate, Egypt, with the predominance of C. hominis genotype in children.

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