HEPATITIS C is an infective disease impacting the liver caused by hepatitis virus Ryan et al 2004 hepatitis degree Celsius virus a member of flaviviridy household was discovered as anew viral agent of non _A non _B, hepatitis virus by choo and coworkers in 1989 choo QC 1989
And is recognized as a major menace to planetary public wellness. ( Lauer 2001 ; Booth 2001 ) . Hepatitis C Virus has positive sense RNA genome that consists of individual unfastened reading frame of 9600 nucleoside bases ( Hwang 2001 ; Muhammad 2005 ) .
At is estimated that 200 million people word broad are infeted with the HCV, the overall prevalence of anti HCV antibodies was 2 % in Spain, 3.9 % in Jakarta, Indonesia and 1.25 % Zakiuthos Greek.
The Seroprvalence of HCV in infirmary based surveies done in Nauritius, Ethiopia of South Indies showed a serprevalance of 5.9 % , 6 % , and 4.8 % severally,
WHO estimates indicate 10.24 million HCV in feted individual in Ind and seroprevalance of 2.4 % in Pakistan.
The seroprevalance of hepatitis “ C ” in different parts of state, reported in last 5 old ages is from2.2 % to 13.5 % the highest serporevalance of hepatitis “ C ” is reported from lahor 13.5 % Jamshero and Mardan 9 % ,
Hpatitis C Virus HCV continues to be amajor disease burdenon the universe. The who estimated prevalence of approximately 3 % with the virus impacting 170 million people worldwide ( universe wellness organisation, 2000 ) . In developing states where resource and installations may be significantly limited the prevalence HCV is higher as compared to the developed universe ( wild and hall, 2000 ) . The prevalence of chronic hepatitis C in the Asia Pacific part is variable between 4 % to 12 % ( Takahashi et al 1993 ) . Approximately 80 % of the persons infected with HCV advancement to chronic infection ( Amarapurkar, 2000 ) .
Hepatitis C virus is transmitted through contaminated blood transfusion, surgery, surgical instruments, dental surgery and inordinate alveolar consonant audiences, sexual contacts, durg maltreatments, sharing of the house hold points such as razors, tooth coppices and shaving from the Barber ( perrillo et Al 1990 ; sato et al 1994 ; van staa 1997 ) . Some wellness attention processs, i.e, surgical and dental interventions, have recentaly been indicated as hazard factors for acute HCV ( Mele and Marzolini, et al 2000 ) . The prevalence of hepatitis C is more than hepatitis B the chief hazard factor is the dental surgery due to internalise instruments, reuse of panpipes and replenishing phials ( Farid et al 2002 ) .
In Pakistan, both HBV pose major hazards as blood borne pathogens ( shah et Al, 2002 ) . Widespread patterns such as insecure injections, improper disposal of risky waste, recycling of used panpipes without proper sterilisation, sharing of acerate leafs by injection drug users and insecure sex are believed to ease the transmittal of these infections, ensuing in high prevalence rates in Pakistan ( Mujeeb, 2001and Khan et Al, 2000 ) . Nosocomial transmittal of HCV is possible when disinfection and sterilisation techniques are unequal, and contaminated equipment is shared among patients. In peculiar, surveies have indicated the possibility of an HCV infection happening among patients on haemodialysis, due to hapless infection control, and the sharing of contaminated medical phials and supplies ( universe wellness organisation, 2000 )
In Pakistan, blood transfusion is still a major beginning of HCV transmittal. Possible grounds for this include deficiency of resources, weak substructure, ill-equipped resources, ill trained staff, inadequate policy execution, frequent power dislocation and morbific showing of blood givers for anti-HCV antibody ( Akhtar S, 2004 ) . Regular blood transfusion in patients withy familial haemolytic anaemia, peculiarly thalassaemia, has improved their overall endurance, but carries a definite hazard of acquisition of blood – borne virus infection, s particularly viral hepatitis ( Alavian et al 2005 ) . Thalassemia patients are peculiarly at hazard of geting HCV by blood transfusion ( Akhtar, 2004 )
Although progresss have been made, a dependable civilization system for HCV is non yet available ( Bendinelli et al 2000 ) . Presently, everyday diagnosing of HCV is based on observing antibodies ( anti-HCV ) in serum by enzyme immunoassay ( EIA ) . However, these methods require venipuncture and expensive equipment, which may non be appropriate for mass sensing proor developing states ( kuo et al, 1989 ) . Other Laboratory checks that are available for the diagnosing and direction of HCV infection include: ( a ) serological trials to observe HCV antibodies ( enzyme linked immunosorbent check ( ELISA ) and recombinant immunoblot check ( RIBA ) , ( B ) molecular trials to observe and quantitate HCV-RNA and ( degree Celsius ) genotyping ( Sandra,2002 )
Aims of the survey are:
To place point beginnings of HCV spread in Mardan.
To analyse nosocomial hazard factors involved with the spread of HCV.
To analyse the efficiency of sterilisation processs and germicides used in the major infirmaries of Mardan.
REVIEW OF LITERATURE
Piazza et Al ( 1995 ) stated that environmental taint of dental surgeries by HCV was investigated by following 35 anti-HCV and HCV-RNA-positive patients with chronic hepatitis through dental intervention ; 328samples were collected from instruments and surfaces after their dental intervention. Twenty ( 6.1 % ) were positive for HCV RNA, including samples from work benches, air turbine handpieces, holders, suction units, forceps, dental mirrors and burs. The writers conclude that, these informations indicate that there is extended taint by HCV of dental surgeries after intervention of Anti-HCV patients and that if sterilisation and disinfection are unequal there is the possible hazard of transmittal to susceptible persons.
Gurevich et Al. ( 1996 ) demonstrates that the possible for perso-to individual transmittal of infective agents such as the human immunodeficiency virus HIV and hepatitis B and C viruses via inadequately sterilized dental instrument exists depending on the prevalence of HIV in the dental patterns. Dental instruments and devices require sterilisation or high-ranking disinfection. An rating of the execution of such procedures was undertaken. Eleven thousand questionnaires on methods used to sterilise and disinfect dental instruments were sent to dental patterns and 1391 ( 13 % ) were retuned for rating. Sixty eight per centum of respondents believed they were sterilising their instruments, nevertheless, some of the liquid chemical merchandises used were non suited for sterilising instruments, and 12 % of respondents used wrong contact times. Forty nine per centum of respondents did non dispute sterilizers with biological spores to look into their map at a
Centers fro Disease Control and Prevention CDC ( 1998 ) published that Nosocomial transmittal of HCV is possible in infection control techniques or disinfection processs are unequal and contaminated equipment is shared among patients. Reports from other states do document nosoceomial HCV transmittal, but such transmittal seldom has been reported in the US, other than in chronic haemodialysis scenes. Surveies indicate that HCV transmittal might happen among patients in a haemodialysis centre because of wrong execution of infection control patterns. It is recommended for all healthcare workers, those who are HCV positive should follow rigorous sterile technique and standard safeguards, including appropriate usage of manus lavation, protective barriers, and attention in the usage and disposal of acerate leafs and other crisp instruments.
Garcia et Al ( 2000 ) stated that proper sterilisation of invasive instruments and equipments is of great importance in the pattern of dental medicine, where assorted blood or saliva borne pathogens e.g. HCV and HBV could be easy transmitted to patients and dental staff via contaminated or inadequately sterilised instruments.
Mujeeb SA et Al ( 2001 ) and Khan AJ et Al ( 2000 ) concluded from their research that widespread patterns such as insecure injections, improper disposal of risky waste, recycling of used panpipes without proper sterilisation, sharing of acerate leafs by shooting drug users and insecure sex are believed to ease the transmittal of these HCV and other infections, ensuing in high prevalence rates in the Pakistan.
BUTT et Al. ( 2003 ) conducted the survey to place the function of dental surgical processs in lending to the transmittal of hepatitis C.78 back-to-back grownup patients of both Sexes enduring signifier chronic hepatitis C Were evaluated. It was concluded that Dental processs were the major beginning of exposure ( 39.7 % ) followed by injections ( 16.6 % ) surgical processs ( 16.6 % ) , diabetes ( 12.8 % ) household history of hepatitis ( 9 % ) blood transfusions ( 7 % ) , tattooing ( 5.1 % ) and history of contact with a icteric patient ( 2.6 % ) . There was a statistically important difference in distribution of hazard factor, with dental processs being the commonest factor ( p & lt ; 0.001 ) . The high prevalence of dental processs in patients with chronic hepatitis C stresses the importance of uneffective infection control methods practiced by dental sawboness as a hazard factor for geting hepatitis C and which were likely the beginning of infection.
Saeed et Al. ( 2004 ) assessed HCV seroprevalence among polytransfused thalasssemic kids registered at a charity clinic and blood bank of a nongovernmental organisation ( NGO ) at Karachi for regular blood transfusion therapy. Between Noveber 1998 and January 1999, back-to-back 256 registered kids were included and a serum sample for rating of anti HCV from each take parting kid was obtained Anti HCV was detected by HCV microparticle enzyme immunoassay 3rd coevals kit harmonizing to the maker ‘s instructions ( Abbott, chicageo, USA ) HCV ribonucleic acid RNA was tested in sera of a subset of anti-HCV positive patients utilizing amplicor HCV RNA assay as suggested by the maker ( Roche diagnostic System, USA ) Eighty nine of 256 ( 34.8 % ) thalassemic kids were anti-HCV positive. HCV RNA was detected in 15 of 38 ( 39.5 % ) anti-HCV positive kids who were tested for HCV RNA. The mean ( A±SD ) age of HCV seropositive kids was 11.9 A± 4.6 old ages.
Khan et Al ( 2007 ) conducted a descriptive survey carried out from July 2003 to July 2004 on 1630 patients admitted in the section of orthopaedicss Ayub Teaching Hospital Abbottabad. Patients of either sex, of all ages undergoing surgery were included in the survey. All patients underwent testing for Hepatitis B and Hepatitis -C and confirmed by Elisa method in positive patients. Out of 1630 patients Hepatitis B and C was present in84 ( 5.15 % ) patients.Out of 84 septic patients 51 ( 3.12 % )
were enduring from hepatitis C and 33 ( 2.02 % ) were enduring from hepatitis B. In 2 ( 0.12 % ) patients both hepatitis B and C infections were present. Among the predisposing gactors old history of surgery was positive in 18 ( 21.43 % ) patients ; history of blood transfusion in 13 ( 15.47 % ) patients, dental process was in 7 ( 8.33 % ) patients, and abroad visit in 4 ( 4.76 % ) patients. It has been concluded that the prevalence of hepatitis B and C in orthopaedic patients is rather high with the common hazard factors.
E Girou et Al. ( 2008 ) behavior ed a prospective observational survey to measure the functions of environmental taint and non-compliance with standard safeguards in cross- transmittal of HCV between patients in a haemodialysis unit. Patients undergoing long term kidney dialysis uncovering 2 instances of HCV transmittal. An probe was so launched to find whether the patients were infected in the haemodialysis unit. Environmental taint by blood and HCV R NA was assessed, as was conformity with recognized infection control safeguards such as manus lavation and usage of baseball mitts. Blood contaminated surfaces may be a beginning of HCV cross transmittal in a haemodialysis unit, “ the survey writers concluded.
MATERIALS AND METHODS
Blood samples will be collected from common population of territory mardan, Samples from peoples of assorted age group will be collected in 1 % PBS solution and transported to labs in Zoology Department Abdul Wali Khan University Mardan for farther analysis.
Peopless who has a recent history of dental surgeries or grading will besides be screened and sampled for the presence of HCV,
Screening for HCV positive IDUs will be carried out with the aid immuno- chromatographic ( accurate ) Blood samples will so be collected from HCV positive persons for farther analysis.
Liver Function Tests ( LFTs )
LFTs such as Alt, AST, Alkaline Phosphatase and hematoidin of ELISA positive samples will be done at Anwar Medical Center and Laboratory Mardan.
HCV RAN Extraction
HCV RNA will be extracted form 100 Aµl serum or samples collected from surgical instruments in PBS either manually or by utilizing Anagen RNA extraction kit ( Purescript, USA ) harmonizing to the maker ‘s direction.
Qualitative HCV RNA sensing
A ) Reverse Transcription PCR
Serum will be prepared in a laminar flow bench and will be frozen at 70C. Qualitative sensing of serum HCV RNA will be performed by RTPCR. Briefly, after extraction of HCV RNA from 100 Aµl serum, complementary DNA will be synthesized utilizing antisense primer at 37 C with M-MLV contrary RNA polymerase enzyme.
a ) Regular PCR
The amplified complementary DNA will farther be subjected to tow unit of ammunitions of PCR elaboration. The status for the first unit of ammunition PCR will be as follows:
An initial denaturation measure at 95C for 2 proceedingss followed by 30 rhythms of 94C for 45 seconds. 54C for 45 secpmds. Amd 72C fpr 1 minute will be performed in Eppendorf thermic cycler ( Eppendorf, Germany ) . The conditions for the 2nd unit of ammunition PCR will be the same except that a different set of interior primers will be used in the PCR mix in order to magnify the 1st unit of ammunition merchandise.
The diagnosing of chronic HCV will be based on elevated serum ALT ( SGPT ) and AST ( SGOT ) degrees, histological scrutiny, and consistent sensing of serum CVHhHhh HCV RNA.
Quantitative step measuring of HCV RNA
Quantification of HCV RNA in pre intervention sera of patients will be performed by RT-PCR with an internal RNA criterion derived from the 5 UT R.
Genotyping of HCV ( harmonizing to the categorization of Simmonds et Al. ( 1993 ) will be performed by in house type-specific PCR at IBGE of Realtime PCR research lab, Peshawar. All taint bar steps suggested BB Kwok and Higuchi ( 1989 ) will be purely followed.
Agarose gel cataphoresis and Gel certification
All the PCR merchandises ( first and 2nd unit of ammunitions ) will be analyzed on 1.8 % agarose gel prepared in 0.5 % TBE buffer, stained with ethedium bromide ( 10 ug/ml ) as florescent dye. Gels will be photographed utilizing alpha quant ( Alpha innotech ) . 100-bp Deoxyribonucleic acid ladder ( Gibco BRL ) will be used as DNA size marker.
The information will be analyzed and the drumhead statistic will be carried out by a statistical bundle, SPSS version 10.0 for Windowss.
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