The prosecution had filed an application in the Islamabad High Court seeking voice samples of the suspects to compare it with the communication intercepted by Indian intelligence and then present it before the anti-terrorism court (ATC) as evidence against the seven suspects in the Mumbai attack case.
The Islamabad High Court on Monday dismissed the petition.
In 2011 and 2015, the issue of obtaining voice samples of Lakhvi had been dismissed by the trial court on the grounds that "no such law exists that allows obtaining of voice sample of an accused".
In the recorded intercepts, the suspects are alleged to be instructing the terrorists.
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The trial court had also rejected another petition of the prosecution requesting the court to declare Ajmal Kasab and Faheem Ansari absconders in order to meet legal formalities.
The prosecution had told the court that unless it declared the two men absconders, the trial against them would remain "inconclusive" as both have been cited as accused in the Mumbai attack case by Indian authorities and that they were also wanted by the Federal Investigation Agency (FIA) that was probing the 26/11 case.
Six accused -- Abdul Wajid, Mazhar Iqbal, Hamad Amin Sadiq, Shahid Jameel Riaz, Jamil Ahmed and Younis Anjum -- have been in Adiala Jail for more than six years in connection with planning and executing the Mumbai attack in November, 2008, that left 166 people killed.
Lakhvi, 56, secured bail in December, 2014, and was subsequently released from Adiala Jail on April 10, 2015, after the Lahore High Court set aside the government's order to detain him under a public security act.