Development of a species-diagnostic polymerase chain reaction assay for the identification of Culex vectors of St. Louis encephalitis virus based on interspecies sequence variation in ribosomal DNA spacers.

نویسندگان

  • M B Crabtree
  • H M Savage
  • B R Miller
چکیده

Culex pipiens complex mosquitoes (Cx. p. pipiens and Cx. p. quinquefasciatus) are among the principal vectors of St. Louis encephalitis (SLE) virus in the eastern United States; Cx. restuans and Cx. salinarius play secondary roles in the transmission and maintenance of the virus cycle. Accurate identification of these three species in field collections is required for epidemiologic studies of SLE virus transmission. We have developed a polymerase chain reaction (PCR) assay for this purpose. Species-specific PCR primers were designed based on interspecies nucleic acid sequence variation in the first and second internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal DNA gene array; however, insufficient variation was detected to differentiate between subspecies of the Cx. pipiens complex. The primers were used together in a single amplification reaction to correctly identify specimens to species using genomic DNA extracted from whole individual mosquitoes, DNA from triturated mosquito pools, or crude DNA from mosquito heads or legs.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

MORPHOLOGY, SYSTEMATICS, EVOLUTION Development of Multiplexed Species Specific Polymerase Chain Reaction Assays for Identification of the Culex (Melanoconion) Species (Diptera: Culicidae) of the Southeastern United States Based on rDNA

Adult female mosquitoes within the subgenus Culex (Melanoconion) Theobald (Diptera: Culicidae) are difÞcult to identify to species using external morphological features. We present two multiplexed polymerase chain reaction assays that quickly and accurately identify specimens from the southeastern United States based on sequence differences in the internal transcribed spacers of the ribosomal D...

متن کامل

Molecular Identification of Vibrio harveyi From Larval Stage of Pacific White Shrimp (Litopenaeus vannamei) Boone (Crustacea:Decapoda)By Polymerase Chain Reaction and 16S rDNA Sequencing

A Gram-negative, rod-shaped, motile and halophilic bacterium identified as Vibrio harveyi was isolated from hatchery-reared larval of Iran Shrimp Research Centerfollowing mass mortality . Seven isolates of Vibrio spp.were collected from nauplii of Litopenaeus vannamei and then were evaluated for characteristics including morphology, physiology and biochemistry. The results indicated that the is...

متن کامل

Molecular Identification of Vibrio harveyi From Larval Stage of Pacific White Shrimp (Litopenaeus vannamei) Boone (Crustacea:Decapoda)By Polymerase Chain Reaction and 16S rDNA Sequencing

A Gram-negative, rod-shaped, motile and halophilic bacterium identified as Vibrio harveyi was isolated from hatchery-reared larval of Iran Shrimp Research Centerfollowing mass mortality . Seven isolates of Vibrio spp.were collected from nauplii of Litopenaeus vannamei and then were evaluated for characteristics including morphology, physiology and biochemistry. The results indicated that the is...

متن کامل

Rapid identification of dengue virus by reverse transcription-polymerase chain reaction using field-deployable instrumentation.

Dengue virus universal and dengue serotype 1 to 4, fluorogenic probe hydrolysis (TaqMan), reverse transcription-polymerase chain reaction assays were developed for screening and serotype identification of infected mosquito vectors and human sera using a field-deployable, fluorometric thermocycler. Dengue universal and dengue 1 to 4 serotype assay in vitro sensitivity and specificity results wer...

متن کامل

Development of New Modified Simple Polymerase Chain Reaction and Real-time Polymerase Chain Reaction for the Identification of Iranian Brucella abortus Strains

Brucellosis is primarily a worldwide zoonotic disease caused by Brucella species. The genus Brucella contains highly infectious species that are classified as biological threat agents. In this regard, the identification of Brucella can be a time-consuming and labor-intensive process posing a real risk of laboratory-acquired infection to the laboratory staff. This stud...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • The American journal of tropical medicine and hygiene

دوره 53 1  شماره 

صفحات  -

تاریخ انتشار 1995