OPTIMIZATION OF REAL-TIME PCR AIMING AT THE MOLECULAR DIAGNOSIS OF TUBERCULOSIS.਀

Thiago Milech de Assunção (INCT-TB); Ana Christina de Oliveira Dias (INCT-TB); Luiz Augusto Basso (INCT-TB); Diógenes Santiago Santos (INCT-TB); Eraldo Luis Batista Jr. (INCT-TB)

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Mycobacterium tuberculosis (Mt) infection remains a serious public health problem due to its high਀爀椀猀欀 漀昀 瀀攀爀猀漀渀ⴀ琀漀ⴀ瀀攀爀猀漀渀 琀爀愀渀猀洀椀猀猀椀漀渀Ⰰ 洀漀爀戀椀搀椀琀礀 愀渀搀 洀漀爀琀愀氀椀琀礀⸀ 䌀甀爀爀攀渀琀氀礀Ⰰ there are approximately 9,2 million new infections and 1,7 million deaths਀愀琀琀爀椀戀甀琀攀搀 琀漀 琀甀戀攀爀挀甀氀漀猀椀猀 ⠀吀䈀⤀ 攀瘀攀爀礀 礀攀愀爀⸀ 倀爀漀最爀攀猀猀椀瘀攀 椀渀挀爀攀愀猀攀猀 椀渀 吀䈀 infections are expected and a worldwide annual incidence of 14,4 million cases਀眀愀猀 瀀爀攀搀椀挀琀攀搀 戀礀 琀栀攀 圀漀爀氀搀 䠀攀愀氀琀栀 伀爀最愀渀椀稀愀琀椀漀渀⸀ 㰀漀㨀瀀㸀㰀⼀漀㨀瀀㸀㰀猀瀀愀渀 猀琀礀氀攀㴀∀∀㸀㰀⼀猀瀀愀渀㸀䐀椀愀最渀漀猀琀椀挀 methods of TB, nowadays, promote an unacceptable਀搀攀氀愀礀 椀渀 搀椀愀最渀漀猀椀猀 愀渀搀 甀渀昀漀爀琀甀渀愀琀攀氀礀 氀攀愀搀 琀漀 攀氀攀瘀愀琀攀搀 昀愀氀猀攀 渀攀最愀琀椀瘀攀 爀攀猀甀氀琀猀⸀㰀⼀猀瀀愀渀㸀 The aforementioned is critical since the infected individuals remain untreated,਀椀渀挀爀攀愀猀椀渀最 琀栀攀 㰀猀瀀愀渀 猀琀礀氀攀㴀∀挀漀氀漀爀㨀 戀氀愀挀欀㬀∀㸀氀椀欀攀氀椀栀漀漀搀 漀昀 搀椀猀攀愀猀攀 琀爀愀渀猀洀椀猀猀椀漀渀 愀渀搀 epidemic aggravation. Thus, the rapid detection and identification of Mt਀椀渀 猀攀挀爀攀琀椀漀渀猀 眀漀甀氀搀 戀攀 栀椀最栀氀礀 搀攀猀椀爀愀戀氀攀 昀漀爀 瀀爀漀瀀攀爀 搀椀愀最渀漀猀椀猀 愀渀搀 攀昀昀攀挀琀椀瘀攀 琀爀攀愀琀洀攀渀琀Ⰰ along with the prevention and control of tuberculosis transmission. Real-time Polymerase਀䌀栀愀椀渀 刀攀愀挀琀椀漀渀 ⠀刀吀ⴀ倀䌀刀⤀ 甀猀椀渀最 瀀爀椀洀攀爀猀 愀渀搀 愀 猀瀀攀挀椀昀椀挀 搀攀琀攀挀琀椀漀渀 瀀爀漀戀攀猀 椀猀 愀 recent approach that offers many advantages such as higher sensitivity,਀猀瀀攀挀椀昀椀挀椀琀礀 愀渀搀 昀愀猀琀 搀椀愀最渀漀猀椀猀⸀ 吀栀攀 愀椀洀猀 漀昀 琀栀攀 瀀爀攀猀攀渀琀 猀琀甀搀礀 眀攀爀攀 琀漀 椀洀瀀氀攀洀攀渀琀 a molecular diagnostic test for tuberculosis, as well as to screen and optimize਀䴀琀 琀愀爀最攀琀 最攀渀攀猀 昀漀爀 刀吀ⴀ倀䌀刀⸀ 㰀猀瀀愀渀 猀琀礀氀攀㴀∀∀㸀㰀⼀猀瀀愀渀㸀匀瀀甀琀甀洀 from patients with diagnosed tuberculosis was collected and processed for DNA਀椀猀漀氀愀琀椀漀渀⸀ 刀吀ⴀ倀䌀刀 眀愀猀 瀀攀爀昀漀爀洀攀搀 漀渀 愀 琀栀攀爀洀愀氀挀礀挀氀攀爀 琀漀 愀洀瀀氀椀昀礀 愀渀搀 搀攀琀攀挀琀 琀栀攀 䴀琀 genes. Primers and probes designed to detect Mt single-copy genes conjugated with਀昀氀甀漀爀漀瀀栀漀爀攀猀 ⠀吀愀焀䴀愀渀⤀⸀ 吀栀攀 挀漀渀猀琀爀甀挀琀椀漀渀 漀昀 猀礀渀琀栀攀琀椀挀 䐀一䄀 猀琀愀渀搀愀爀搀猀 昀漀爀 䴀琀 猀攀爀瘀攀搀 as a references for absolute quantification of nucleic acids, which reflected਀愀戀猀漀氀甀琀攀 渀甀洀戀攀爀猀 漀昀 䴀琀⸀ 吀栀漀猀攀 眀攀爀攀 漀戀琀愀椀渀攀搀 昀爀漀洀 愀 瀀爀椀漀爀 愀洀瀀氀椀昀椀挀愀琀椀漀渀 愀渀搀 cloning of genomic DNA of Mt, targeting the intergenic sequences as noted above.਀吀栀攀 爀攀猀甀氀琀猀 猀栀漀眀攀搀 琀栀愀琀 䴀礀挀漀戀愀挀琀攀爀椀甀洀 䐀一䄀 眀愀猀 搀攀琀攀挀琀攀搀 椀渀 䐀一䄀 椀猀漀氀愀琀攀搀 昀爀漀洀 sputum; nevertheless, control individuals failed to show any signs of fluorescence਀椀渀挀爀攀愀猀攀Ⰰ 挀漀爀爀漀戀漀爀愀琀椀渀最 琀栀攀 愀戀猀攀渀挀攀 漀昀 挀爀漀猀猀 愀洀瀀氀椀昀椀挀愀琀椀漀渀 漀昀 栀漀猀琀 䐀一䄀 愀猀 眀攀氀氀 as DNA from other pathogens recovered from the patients. The positive results਀漀戀琀愀椀渀攀搀 挀愀氀氀 昀漀爀 昀甀爀琀栀攀爀 攀砀瀀攀爀椀洀攀渀琀猀 愀椀洀椀渀最 愀琀 搀椀愀最渀漀猀椀猀 漀昀 瀀愀甀挀椀戀愀挀椀氀愀爀 吀䈀⸀㰀漀㨀瀀㸀㰀⼀漀㨀瀀㸀㰀⼀猀瀀愀渀㸀㰀⼀瀀㸀 ਀ ਀㰀⼀昀漀渀琀㸀㰀⼀瀀㸀㰀戀爀㸀㰀戀㸀倀愀氀愀瘀爀愀猀ⴀ挀栀愀瘀攀㨀 㰀⼀戀㸀☀渀戀猀瀀㬀搀椀愀最渀漀猀琀椀挀Ⰰ 䴀礀挀漀戀愀挀琀攀爀椀甀洀 琀甀戀攀爀挀甀氀漀猀椀猀Ⰰ 爀攀愀氀ⴀ琀椀洀攀 倀䌀刀㰀⼀琀搀㸀㰀⼀琀爀㸀㰀⼀琀愀戀氀攀㸀㰀⼀琀爀㸀㰀⼀琀搀㸀㰀⼀琀愀戀氀攀㸀㰀⼀戀漀搀礀㸀㰀⼀栀琀洀氀㸀